International Journal Of Cancer Research Articles

Abstract A115: The CD95 ligand inhibitor APG 101 reduces tumor recurrence and metastasis in an adjuvant orthotopic mouse model of pancreatic cancer as well as primary tumor load in a palliative setting

Abstract Since the first description of CD95/Fas this member of the death receptor superfamily has been regarded as paradigmatic for death receptors, capable of induction of caspase-driven apoptosis. More recently, CD95 has also emerged as an inducer of other signaling pathways including pro-inflammatory and migratory mechanisms (for an overview see Roeder C. et al., Eur J Cell Biol. 2011). Previously, we demonstrated a strong induction of NFkB activity after CD95 stimulation of pancreatic cancer cells resulting in increased invasiveness under in vitro conditions. Inhibition of CD95 mediated mechanisms in tumor gains importance since malignant cells may misuse this system for progression as demonstrated by others eg. for ovarian cancer and glioblastoma. From a clinical point of view, several observations also point into the same direction. Studies on colorectal cancer, head and neck, hepatocellular carcinoma for example suggest CD95 ligand expression to correlate negatively with patients prognosis. APG101 is an inhibitor of CD95 ligand consisting of the CD95 receptor extracellular domain fused to the Fc domain of IgG. A randomized controlled phase II ‘proof-of-concept’ study in patients with recurrent glioblastoma was successfully completed recently (NCT01071837). Up to date there exist no in vivo studies concerning the influence of blocking the interaction between CD95 and its ligand using this recombinant inhibitor in pancreatic cancer. Thus we tested this novel tool in a palliative and adjuvant orthotopic xenotransplantation model of human pancreatic cancer. In this study PancTuI Luc cells were used, which establish a highly infiltrating primary tumor with very low/no metastatic capacity in the liver. All mice (n=8/group) survived the operative procedures for the palliative treatment and were randomly assigned into five groups: control group; treatment with different concentrations of APG101: 12.5, 25, 50, and 100 µg/g bw. (i.p. injection, twice per week). Upon tumor resection (2/3rd pancreatectomy 10 days after tumor cell inoculation) local as well as distant recurrent disease (primarily in the liver) is regularly observed. All mice surviving the operative procedures for the adjuvant treatment (n=9/group) were randomly assigned into two groups: control group with 200 µl saline 0.9 % i.p. and the APG 101 group with a dose of 25 µg/g bw i.p (twice a week). Treatment was started three days after subtotal pancreatectomy and lasted for 28 days until sacrifice of the animals. Palliative treatment: mean tumor weight was 353 ± 179 mg for animals in the control group compared to 210.4 ± 86.5 mg to the mice treated with 25 µg APG101/g bw (p = 0.09) and 221.4 ± 94 mg (p = 0.021) in the group treated with 50 µg APG101/g bw Tumor volume was significantly reduced in all treatment groups, again with 25 µg APG101/g bw achieving maximal growth inhibition. Within the 4 weeks no metastases occurred in any group as previously observed within this model. Adjuvant treatment: All mice developed recurrent tumors. The mean weight of locally recurrent tumor was 887.5 ± 467.3 mg in the control group (treated with 0.9% saline). The other group treated with APG101 (25 µg/g bw) showed a local tumor recurrence with a significant lower tumor weight (p=0.027). The average number of metastases per mouse in the therapy group was 2.3 versus 4 metastases / mouse in the control group. Reduced tumor growth in both therapeutic settings was reflected by a reduced Ki67 proliferation index. Our data underline the importance of the CD95 antagonist APG101 as a new option in pancreatic cancer therapy that might be further evaluated also in combination with chemotherapies. Note: These results were published in the International Journal of Cancer as a full-length article before the online publication of this abstract. Citation information: Inhibition of IL-6 signaling significantly reduces primary tumor growth and recurrencies in orthotopic xenograft models of pancreatic cancer. Goumas FA, Holmer R, Egberts JH, Gontarewicz A, Heneweer C, Geisen U, Hauser C, Mende MM, Legler K, Röcken C, Becker T, Waetzig GH, Rose-John S, Kalthoff H. Int J Cancer. 2015 Jan 21. doi: 10.1002/ijc.29445. [Epub ahead of print] PMID: 25604508. Citation Format: Freya Goumas, Khalid Rhashid, Anna Trauzold, Harald Fricke, Michael Kluge, Thomas Becker, Jan Egberts, Holger Kalthoff. The CD95 ligand inhibitor APG 101 reduces tumor recurrence and metastasis in an adjuvant orthotopic mouse model of pancreatic cancer as well as primary tumor load in a palliative setting. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Innovations in Research and Treatment; May 18-21, 2014; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2015;75(13 Suppl):Abstract nr A115.

Expression of the axon guidance protein Robo1 in pancreatic ductal adenocarcinoma from smokers compared to nonsmokers.

305 Background: The most important modifiable risk factor for the development of pancreas cancer is smoking, which accounts for up to 25% of pancreatic ductal adenocarcinomas (PDAC; Maisonneuve P, Lowenfels AB: Epidemiology of pancreatic cancer: an update. Digestive Diseases 28:645-656, 2010), and the incidence of PDAC correlates with smoking prevalence (Weiss W, Benarde MA: The temporal relation between cigarette smoking and pancreatic cancer. American journal of public health 73:1403-1404, 1983). A recently published Ingenuity Pathway Analysis of GWAS genotype and risk factor data from the Pancreatic Cancer Case Control Consortium demonstrated that axon guidance signalling genes were significantly overrepresented in smokers (Tang H, Wei P, Duell EJ, et al: Axonal guidance signaling pathway interacting with smoking in modifying the risk of pancreatic cancer: A gene and pathway-based interaction analysis of GWAS data. Carcinogenesis:bgu010, 2014) and we aimed to investigate this link in our cohort. Methods: Tissue from resected PDAC were obtained from 200 patients via the Australian Pancreatic Cancer Genome Initiative with patient consent and ethical approval. Immunohistochemistry was performed on TMAs with the anti-Robo1 antibody (ab7279). The sections were scored on a 4-point scale of 0, 1, 2 and 3 intensity. All sections were scored blindly by two independent reviewers. Results: Robo1 protein expression is found in the normal pancreas, predominantly in acinar cells. In PDAC, Robo1 expression is predominantly in epithelial ductal cells. Most PDACs have Robo1 expression with an overall mean score of 1.65±0.05. 20 out of 200 patients were current smokers at the time of their pancreatectomy, 97 were never smokers, the remainder were ex-smokers. The mean score for smokers was 2.1± 0.1 and for never smokers 1.6±0.1 (p = 0.0003). Genomic analysis did not demonstrate any mutations in the Robo1 gene. There were 7 cases of loss of heterozygosity for Robo1; none of these were current smokers and they had an average score of 1.25±0.17. Conclusions: In addition to our genomic (Biankin AV, Waddell N, Kassahn KS, et al: Pancreatic cancer genomes reveal aberrations in axon guidance pathway genes. Nature 491:399-405, 2012) and methylation data (Nones K, Waddell N, Song S, et al: Genome‐wide DNA methylation patterns in pancreatic ductal adenocarcinoma reveal epigenetic deregulation of SLIT‐ROBO, ITGA2 and MET signaling. International Journal of Cancer, 2014), we provide the results of protein expression of Robo1 in a clinically annotated cohort of 200 cases of PDAC. We demonstrate that patients who are currently smoking have enhanced Robo1 expression. Preliminary results indicate that this may confer a poorer prognosis when coupled with high SLIT2 expression.

International Journal of Cancer and Clinical Research- Commited to Ensure Quality Publication in Cancer Research

C l i n M e d International Library Citation: Mohammed SI (2014) International Journal of Cancer and Clinical ResearchCommited to Ensure Quality Publication in Cancer Research. Int J Cancer Clin Res 1:001e Received: August 14, 2014: Accepted: August 18, 2014: Published: August 20, 2014 Copyright: © 2014 Mohammed SI. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Mohammed, Int J Cancer Clin Res 2014, 1:1

Abstract SY37-03: Lifting the metabolic break on caspase 2 to enhance chemotherapy

Abstract Chemotherapeutic elimination of many tumors depends on the induction of programmed cell death (apoptosis) in tumor cells. We and others have found that DNA damaging chemotherapeutics (e.g., cisplatin) and cytotoxic agents (e.g., paclitaxel) commonly used in cancer therapy (for eg. ovarian cancer, breast cancer and others) requires the initiator caspase, caspase 2, for robust induction of apoptosis. However, resistance to chemotherapy-induced apoptosis remains a significant obstacle in the treatment of these cancers. Recently, it has been recognized that the increased metabolic rate found in many cancer cells is a contributing factor in chemoresistance. While the mechanism through which metabolism regulates chemoresistance is not fully elucidated, we have found that caspase 2 activity is subject to metabolic control. Specifically, a highly active pentose phosphate pathway (PPP) leads to high levels of NADPH. This promotes suppressive phosphorylation of caspase 2, thereby inhibiting cancer cell death. Glucose-6-phosphate dehydrogenase (G6PD) is the rate-limiting enzyme for entry of G6P into the PPP. G6PD has been reported to promote oncogenic transformation in vitro and in vivo and G6PD activity is elevated in a variety of cancers (1). Moreover, loss of G6PD in both Chinese hamster ovary cells and human fibroblasts has been reported to enhance radiosensitivity (2, 3). PPP flux is enhanced in cancer cells through a variety of mechanisms: it has been reported that mutation of the tumor suppressor p53 prevents its suppression of G6PD and overexpression of TAp73 increases G6PD expression in tumors (4). Our data suggest that a highly significant consequence of increased G6PD levels is the suppression of caspase 2. This is independent of any effects G6PD may have on redox potential in tumors. We now show that lifting this suppression of caspase 2, through inhibition of G6PD, will markedly sensitize ovarian, breast and pancreatic cancer cells to chemotherapeutics. G6PD is likely to be a good drug target for cancer therapy: loss of G6PD is known to have minimal adverse consequences for humans, with nearly 7.5% of the world's population exhibiting some degree of G6PD enzyme deficiency (5). Thus, inhibition of G6PD in normal cells (particularly in the temporally restricted setting of cancer therapy) is likely to cause little toxicity. On a mechanistic level, we have reported that the metabolic control of caspase 2 is exerted via its inhibitory phosphorylation by activated CaMKII (6). We now demonstrate a novel form of control of CaMKII activation by metabolism that depends, at least in part, on dephosphorylation of CaMKII at previously uncharacterized sites (T393/S395). We show that this dephosphorylation is mediated by metabolic activation of protein phosphatase 2A (PP2A) in complex with the B55β targeting subunit and that metabolic activity controls the association of the B55β/PP2A complex . This represents a novel locus of CaMKII control and also provides a mechanism contributing to metabolic control of apoptosis. These findings may also have implications for metabolic control of the many CaMKII-controlled and PP2A-regulated physiological processes, as both enzymes appear to be responsive to alterations in glucose metabolized via the PPP. Finally, we will discuss our recent findings involving a role for caspase 2 in the induction of lipoapoptosis in the setting of fatty liver disease, the most severe form of which (nonalcoholic steatohepatitis) may predispose patients to development of liver cancer. We have found that fatty liver can promote a dramatic induction of caspase 2 in both mouse models of disease and in human patients and will present data to suggest that this is a critical component of disease etiology.

Abstract B23: P1 glycosphingolipid is expressed on ovarian cancer cells recognized by naturally occurring anti-P1 antibodies

Abstract Using printed glycan array we have recently demonstrated that natural occurring anti-glycan antibodies to chemically synthesized P1 trisaccharide and Pk are significantly reduced in ovarian cancer patients (1). These findings were successfully validated with two in-house developed independent glycan-based immunoassays (ELISA and suspension array) (2). Both candidates are glycosphingolipids (GSL) and share similar structural motifs. P1 naturally occurs as a pentasaccharide belonging to the neolacto series. Here we aim to a) validate our findings in an independent validation cohort, b) identify the immunoglobulin subtype causing this observation, c) identify the corresponding naturally expressed P1 antigen in cancer cells, and c) purify human anti-P1 antibodies from cancer ascites. A cohort of 155 blood plasma samples comprising healthy controls and patients with cancer of the ovary, tube and peritoneum was profiled for P1 antibodies using monoplex suspension array. GSL isolated from fresh cancer tissue were PVDF blotted and extracted enzymatically following purification of glycan alditols. Glycan structures were identified by negative-mode LC-ESI-MS. P1-expressing cancer cell lines were detected using flow cytometry. Human anti-P1 antibodies were affinity purified from cancer ascites and studied using glycan-based immunoassays. Glycan-based immunoassay profiling of validation cohort confirmed our recent findings, reduced anti-P1 antibody levels in cancer patients compared to healthy controls (p=0.0002). We could also demonstrate that this observation was caused by IgM subtypes and not IgG. Corresponding P blood group- related antigens (GSL) including P1 and Pk were identified in cancer tissue samples. Flow cytometry revealed that a serous ovarian cancer cell line is positive for P1 (up to 46%). We further demonstrate that affinity purified anti-P1 antibodies bind naturally expressed P1 GSL on ovarian cancer cell surface. Monoclonal as well as affinity purified anti-P1 antibodies applied to identify P1-expressing cancer cell lines revealed high specificity to P1 shown by inhibition assay and glycan-based immunoassays. In this study we demonstrated anti-P1 IgM levels are decreased in a significant number of cancer patients and that the GSL P1 is present on ovarian/ peritoneal cancer cells, both in tissue and immortalized cell lines. We can therefore conclude that P1 is an epitope for human circulating anti-glycan antibodies. These results are in concordance with our previous findings and support the presence of a new antigenic tumor-associated GSL in ovarian cancer. The study of the molecular profile leading to the presence of P1 in ovarian cancer cells is presently under investigation. 1. Jacob F, et al. (2012) Serum antiglycan antibody detection of nonmucinous ovarian cancers by using a printed glycan array. International Journal of Cancer 130(1):138-146. 2. Pochechueva T, et al. (2011) Comparison of printed glycan array, suspension array and ELISA in the detection of human anti-glycan antibodies. Glycoconjugate journal 28(8-9):507-517. Citation Format: Francis Jacob, Merrina Anugraham, Alam Shahidul, Nicolai Bovin, Nicole Packer, Neville Hacker, Viola Heinzelmann-Schwarz. P1 glycosphingolipid is expressed on ovarian cancer cells recognized by naturally occurring anti-P1 antibodies. [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 B23.

Erratum to “Ferritin in Adult-Onset Still’s Disease: Just a Useful Innocent Bystander?”

There is an error in the citations of our previous paper as reference [51] G. Zandman-Goddard, M. Blank, P. Langevitz et al., “Antiserum amyloid component P antibodies in patients with systemic lupus erythematosus correlate with disease activity,” Annals of the Rheumatic Diseases, vol. 64, no. 12, pp. 1698–1702, 2005 should be replaced by the following references: [1] Li, X., et al., “Epidermal Growth Factor-Ferritin H-Chain Protein Nanoparticles for Tumor Active Targeting,” Small, 2012; [2] Jezequel, P., et al., “Validation of tumor-associated macrophage ferritin light chain as a prognostic biomarker in node-negative breast cancer tumors: A multicentric 2004 national PHRC study,” International Journal of Cancer, 2012. 131(2): p. 426–37. [3] Pitcher, J., et al., “Disruption of neuronal CXCR4 function by opioids: preliminary evidence of ferritin heavy chain as a potential etiological agent in neuroAIDS,” Journal of Neuroimmunology, 2010. 224(1-2): p. 66–71. [4] Rosen, H. R., “Placental isoferritin-associated p43 in pregnancy and breast cancer: minireview,” Neoplasma, 1996. 43(6): p. 357–62.

Abstract 67: Characterization of the role of KLF9 transcription factor in breast cancer estrogen response

Abstract Kruppel-like factor 9 (KLF9) is a member of the KLF family of transcription factors. Previous studies report significant KLF member involvement in breast cancer estrogen response, notably KLF4 [1, 2] and KLF5 [3, 4]. KLF9 has been shown in uterine cancer cells to function as an estrogen-dependent regulator of the estrogen response pathway [5]. We aim to evaluate the role of KLF9 in breast cancer cell estrogen response. We have performed studies demonstrating that KLF9 exhibits an early-activated estrogen response. Five sites have been identified upstream of the KLF9 gene that interact with estrogen receptor alpha (ERα) [6-8]; we observe ERα enrichment at three of these sites that is estradiol-dependent. Though KLF9 has been shown to act as an essential element in estrogen response in the uterus, KLF9 response in breast cancer cells has yet to be characterized. To study the role of KLF9 in estrogen-mediated responses in transcriptional and proliferative activity, we have manipulated the level of KLF9 expression in MCF-7 breast cancer cells. These cells have been shown to exhibit a clear transcriptional response to estradiol in luciferase reporter assays. We have optimized RNAi conditions to achieve significant knockdown of the KLF9 gene in MCF-7 cells. Additionally, we have produced and tested a KLF9 overexpression vector construct that significantly upregulates expression of KLF9 in this cell line. These tools will be used to more extensively characterize the role of KLF9 in MCF7 cell estrogen-stimulated transcription and proliferation. We are measuring transcriptional response to estrogen signaling with luciferase reporter assays and estrogen stimulated cell proliferation with BrdU proliferation assays. Elucidating KLF9 involvement in E2-mediated breast cancer cell signaling and response, therefore, is an important component of understanding of the regulatory mechanisms behind estrogen response in breast cancer. 1. Akaogi K et al. (2009). Oncogene 28(32): 2894. 2. Quintana AM et al. (2011). BMC Cancer 11: 30. 3. Guo P et al. (2010). International Journal of Cancer 126(1): 81. 4. Zhao KW et al. (2011). Biochemical Journal 437(2): 323. 5. Velarde MC et al. (2007). Molecular Endocrinology 21(12): 2988. 6. Carroll JS et al. (2006). Nature Genetics 38(11): 1289. 7. Hua S et al. (2008). Molecular Systems Biology 4:188. 8. Fullwood MJ et al. (2009). Nature 462(7269): 58. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 67. doi:1538-7445.AM2012-67

Retraction

British Journal of HaematologyVolume 152, Issue 6 p. 788-788 Free Access Retraction First published: 25 February 2011 https://doi.org/10.1111/j.1365-2141.2011.08572.xAboutSectionsPDF 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 onFacebookTwitterLinked InRedditWechat The following article from the British Journal of Haematology, ‘A modified version of galectin-9 induces cell cycle arrest and apoptosis of Burkitt and Hodgkin lymphoma cells’ by Shoko Makishi, Taeko Okudaira, Chie Ishikawa, Shigeki Sawada, Toshiki Watanabe, Mitsuomi Hirashima, Hajime Sunakawa and Naoki Mori, published online on 22 May 2008 in Wiley Online Library (http://www.wileyonlinelibrary.com) has been retracted by agreement between the authors, the journal Editor in Chief, Finbarr Cotter and Blackwell Publishing Ltd. The retraction has been agreed due to the inclusion in the article of images and data previously published in Blood, 2010;115: 2220-30, Infection & Immunity, 2009; 77: 1337-48, Cancer Science, 2008; 99: 2286-94, Infection & Immunity, 2007; 75: 5223-32, International Journal of Cancer, 2007; 120: 2251-61 and International Journal of Cancer, 2007; 120: 1811-20. The corresponding author, Naoki Mori, takes full responsibility for the duplication of figures in this paper, and states that none of the co-authors were involved in or aware of these events. Reference Makishi, S., Okudaira, T., Ishikawa, C., Sawada, S., Watanabe, T., Hirashima, M., Sunakawa, H. & Mori, N. (2008) A modified version of galectin-9 induces cell cycle arrest and apoptosis of Burkitt and Hodgkin lymphoma cells. British Journal of Haematology, 142, 583– 594. Wiley Online LibraryCASPubMedWeb of Science®Google Scholar Volume152, Issue6March 2011Pages 788-788 ReferencesRelatedInformation

No increased incidence for GB‐virus C infection in a cohort of HIV‐positive lymphoma patients

Dear Sir, Until recently no pathogenic potential could be demonstrated in humans infected with GB-Virus C (GBV-C). In contrast, replicative GBV-C infection in HIV-infected individuals correlated with reduced progression to immunodeficiency (Tillmann, NEJM 2001). We therefore read with great interest, the article by Krajden et al. published recently in the International Journal of Cancer, in which the authors described a cohort of 553 patients with Non-Hodgkin lymphoma and 438 controls for evidence of serum GBV-C RNA. GBV-C RNA was found in 4.5% of NHL patients and in 1.8% of controls, a difference which under analysis achieved statistical significance (odds ratio 1⁄4 2.72, confidence interval1⁄4 1.22–6.69). This study however exclusively included apparently immunocompetent patients and therefore does not offer insight into the potential role of replicative GBV-C infection contributing to the preexisting increased risk of lymphoma in HIVinfected patients. To further clarify the prevalence of GBV-C infection in our cohort of HIV-infected patients with and without lymphoma, we performed a retrospective analysis. In total 33 HIV-positive patients with lymphoma were recruited from the HIV-outpatient clinic at Hanover Medical University for GBV-C RNA and the frequency of viremia was compared with 584 HIV-infected patients without lymphoma. From the 33 patients in the lymphoma group with a mean CD4þ T cell count of 371/lL (range 7–970 cells/lL) 7 had Hodgkin Lymphoma (HL) and 26 Non-Hodgkin Lymphoma (NHL). In total ten patients (30.3%) tested GBV-CRNA positive. In our control group 138 of 584 HIV-positive patients tested positive for GBV-C-RNA (23.6%). These results however failed to achieve statistical significance regarding an association between positive GBV-C-Status and lymphoma (p 1⁄4 0.39). Interestingly the updated results in our cohort revealed an increase in the overall incidence of GBV-C viremia from 16.8% (Tillmann et al. 2001) to 23.6%. This might indicate a general spread of GBV-C infection in our cohort with known elevated risks for parenteral or mucosal transmission of viral infections. Hence the elevated GBV-C incidence in lymphoma patients, described by Krajden et al. could be explained and a confounding phenomenon indicating elevated risks for transmission of other hypothetically oncogenic viruses. On the other hand, the persistence of GBV-C viremia in HIV-positive individuals has been explained by the prevalence of T-cellular immunodeficiency. Therefore it cannot be ruled out that an elevated incidence of GBV-C replication in lymphoma patients rather indicates a secondary or preexisting alteration of T-cell immunity than a causal factor for tumor ontogenesis. Yours sincerely, Diana Ernst Sven Pischke Mark Greer Heiner Wedemeyer Matthias Stoll

Biospecimen use in cancer research over two decades.

Demand for biospecimens in cancer research has increased but there are relatively few data on the trends in biospecimen usage. These data are needed to enable projection of future demand. We analyzed biospecimen usage in publications published at five-year intervals (2008, 2003, 1998, 1993, and 1988) in four cancer research journals (Cancer Research, Clinical Cancer Research, British Journal of Cancer and International Journal of Cancer). We categorized publications in three ways: 1) biospecimen utilization yes/no; 2) biospecimen cohort size; and 3) format of biospecimens used including frozen tissue, Formalin-Fixed Paraffin-Embedded (FFPE) tissue, fresh tissue, fluids, and hematological biospecimens. Biospecimens were used in 1292/3307 (39%) of publications analyzed and sufficient information was available to further classify biospecimen usage in 1228 publications. The proportion of publications in each journal using biospecimens ranged from 23% to 61% between journals, with no significant change within each journal over time. A more detailed review of tissue biospecimen use showed a significant increase in cohort sizes from 1988 to 2008 (mean 52 to 198, respectively; P < 0.0001). This reflected increased cohort sizes for both frozen and FFPE tissues from 1993 to 2008 (frozen, 59 to 119; FFPE, 66 to 194) but not fresh tissues. The relative proportion of studies using frozen or fresh tissues alone has decreased (71% to 24%) while those using FFPE alone or combined FFPE/frozen tissue cohorts has increased (24% to 72%) over this period. We conclude that the overall demand for biospecimens in cancer research has increased significantly (almost fourfold) over the past 20 years. We predict that average cohort sizes will increase by at least twofold for frozen and FFPE biospecimens over the next ten years, and that the majority of studies will be based on FFPE tissues or combined FFPE/Frozen tissue cohorts.

Obligation for cell line authentication: Appeal for concerted action

Cell lines isolated from various human cancers are widely used to study the mechanisms of tumor development and as preclinical models for tumor diagnosis and therapy. This key role played by cell lines in basic and applied cancer research is often taken for granted, but the chronic problem of cell line crosscontamination as well as of altered function due to over-subculturing—first recognized for HeLa cells more than three decades ago—has now become a major issue. As stated in a recent editorial published in Nature 1 ‘‘many studies have found that 18 to 36% of cell lines contain misidentified cells,’’ data which have been confirmed by the major cell repositories. Despite several recent initiatives to correct this intolerable situation, such as the open letter from a panel of scientists led by Roland Nardone to the US Secretary of Health and Human Services in 2007 2 and several subsequently published research articles, reviews and letters all confirming the dilemma of a still growing use of cross-contaminated cell lines, little has changed in reality. Many papers are still being submitted to this and other journals using mis-identified cell lines and many authors seem to be unaware of the problem. This situation prompted the Editors of International Journal of Cancer (IJC) to take the initiative and to impose strict rules for papers submitted for publication in the IJC beginning with the year 2010. In our new Instructions for Authors we now require that ... ‘‘manuscripts dealing with established cell lines have to provide authentication of cells by DNA profiling using Short Tandem Repeats (STR).’’ This ‘‘simple solution (of) routine, cheap DNA profiling’’—as stated in the Nature editorial—can be conducted at each laboratory using commercially available kits and is provided as a service by most cell culture repositories at a reasonable cost (see respective link in the Instructions for Authors). One major prerequisite for a reliable and comparable DNA profiling according to Nature—: ‘‘an electronic database of authenticated DNA profiles’’—of known cell lines has now been established by a consortium, thanks to the initiative and persistent activity of Willy Dirks of the German Cell Bank (DSMZ). This consortium, including major cell banks, has compiled a reference database of more than 2,300 known cell lines that is accessible online and with which scientists can compare their own DNA profiling data for authenticating a given cell line (see Dirks et al. Letter to the Editor, this issue, and the respective link to the database). Moreover, due to the long-standing efforts of Ian Freshney, a pioneer in cell culture technology and advocate of cell line authentication, a list of already confirmed cross-contaminated and mis-identified cell lines with the relevant references has been compiled; this is also freely accessible (see Freshney, Letter to the Editor, this issue, and respective link in the letter and Instructions for Authors of IJC). With these tools authors can easily see whether the cell line they want to use is already ‘indexed’ as cross-contaminated or mis-identified before they start with further authentication procedures. Thus, a solid basis has now been established to eliminate mis-identified cell lines that have produced misleading results and wasted funding, provided the scientific community will accept this offer. One prerequisite for the general acceptance of such a regulation is a concerted action of scientific journal editors and funding agencies to reject all manuscripts and grant applications (based on cell line data) without proper cell line authentication. When we first published a Mini Review in IJC in January 2008, we had addressed a Letter to the Editors-in-Chief of several leading cancer and cell biology journals asking them to join us in this effort, but got little to no response. Therefore, we now reiterate our appeal to the Editors-in-Chief of all life science journals publishing manuscripts with data based on cell lines to require proper authentication of all cell lines. This same petition is also being extended to all granting organizations to accept only grant applications with authenticated cell lines. A letter to major granting agencies has been sent by the Editors of IJC also requesting them to allow additional funds for cell line authentication. Some journals have already included respective demands in their instructions and others have indicated their willingness to do so, but too little has actually happened. The Editors of IJC are convinced that the time is overripe now to take consequent, concerted action on the part of all life science journals and granting agencies and to take the chance to defeat this ‘‘chronic disease’’ in cell culture research.

KAI1/CD82 is a novel target of estrogen receptor-mediated gene repression and downregulated in primary human breast cancer (Int J Cancer 123: 2239-2246, 2008)

With interest I read the article by Christgen et al. that was recently published under the Fast Track heading in the International Journal of Cancer.1 The major reported findings, nicely summarized in the article's title, are that (a) KAI1/CD82 is downregulated in 77% of human breast carcinomas and (b) KAI1 downregulation is strongly correlated with ER positivity. I do not question the accuracy of the reported data. However, I take exception to the authors' claim, repeated several times throughout their article, that their major findings are novel. The fact is that my colleagues and I published practically identical observations 3 years ago.2 Specifically, we reported downregulation of KAI1 expression in 74 of 98 (76%) archival breast carcinomas, and we mentioned that KAI1 downregulation was significantly more common in ER-positive (48 of 55, 87%) versus ER-negative (22 of 36, 61%) tumors (p = 0.0052). Although the focus of our manuscript was on new cell membrane biomarkers involved in breast oncogenesis, we very clearly detailed our observations on KAI1 expression in human breast cancer tissues and 9 cell lines including MCF-7, which Christgen used as a negative control. We published our data in Clinical Cancer Research, which can hardly be considered an obscure journal. It is hard to imagine that even a cursory review of the literature involving KAI1/CD82 and breast cancer would not have brought our article to Christgen et al.'s attention. A scientific finding does not become novel by ignoring its previous description by other investigators. It would have been appropriate for Christgen etal. to acknowledge our published data. Of course, in so doing they would have had to admit that most of their data are confirmatory rather than novel, notwithstanding the fact that they documented the inverse relationship between ER and KAI1 expression in greater detail. Christgen et al. also give the impression that their immunohistochemical staining method for KAI1 is a major technological advance. In fact, they used the same anti-KAI1 antibody and the same heat induced epitope retrieval method that we published in 2005.2 Already in 1999, we had demonstrated that KAI1 protein expression can reliably be evaluated in fixed tissues by immunohistochemistry using a different antibody.3 Christgen also confirms our original observation that KAI1 expression is not correlated with axillary lymph node involvement, tumor grade, or c-erbB-2 status.2 Christgen et al. describe several in vitro experiments as well. They show that the growth and migration of ER-positive, but not ER-negative, breast cancer cells is inhibited by fulvestrant, an antiestrogenic agent, and that this is correlated with up- or downregulation of 4 indicator genes. The authors acknowledge that those experiments confirm previously published observations by other research groups.4-7 It seems that Christgen et al.'s only original and novel contribution is the finding that KAI1 is upregulated by fulvestrant in 2 ER-positive breast cancer cell lines. The International Journal of Cancer may consider following the example of other scientific journals that discourage authors from inserting superlatives (“first”, “largest”, etc.) into their manuscripts. Some of those claims do not stand up to scrutiny. Evidently the reviewers assigned to Christgen's article were not sufficiently familiar with the literature to rebut the authors' exaggerated claims of novelty. In conclusion, I believe the data presented by Christgen et al. are interesting and a useful addition to the scientific literature. However, their major findings are confirmatory (rather than novel) in nature. It is inappropriate for the authors to suggest that their methodology and experimental results are groundbreaking, when they essentially replicate portions of an article that they unfortunately chose not to cite. Yours sincerely, Joseph Geradts MD

Editorial

May we take the opportunity in the first issue of the New Year to express thanks to all our readers, authors, reviewers and editorial board members for their continued interest and help with our journal. Scientific peer review has become a fast-moving undertaking at the International Journal of Cancer since we introduced the online submission and peer review procedures in 2003. Since then, our review times have dropped by nearly 38%, and we can presently boast of average times to decision of about 24 days. The next logical step, then, was to reduce the times for accepted papers to appear online and be made available to our authors and readers. To this end, a new feature was launched on our homepage (http://www3.interscience.wiley.com/journal/29331/home) last August. Perhaps, you have already even noticed the new tab “Accepted Articles.” There, accepted IJC articles in their final form—including the DOI—are posted in pdf format on our Wiley Interscience site 5 days after the publisher receives them. This means that the articles are available to all and can even be cited by DOI while they are being processed. After proofing and approval for publication, the published articles are automatically moved over to “Early View” and then on to their final destination in a print issue. You might want to watch out for the second innovation, too. In the near future, our readership can look forward to reading the “Editor's Choice.” Selected articles of high interest will be posted online and made available free of charge. Through this feature, the International Journal of Cancer can do its share in ensuring that the most important findings published in the journal are available to scientists worldwide, with no strings attached. To accompany this progress, our homepage will also soon be receiving a new look as Wiley-Blackwell will be launching its new web presentation later this year. You may want to keep an eye out for this as we can all expect a wide range of new internet options and advantages. Indeed, we are very grateful that our publisher, Wiley-Blackwell, and the UICC have promoted and implemented these innovations and our special thanks go to them. In 2009, the International Journal of Cancer will continue to pursue the goal of publishing articles of high scientific quality. We look forward to working with our authors, reviewers and readers to this end. The Associate Editors of the IJC join me in wishing you all a Happy and Successful New Year.

Comment on “Production of multivalent protein binders using a self‐trimerization collagen‐like peptide scaffold”

The recently published manuscript by Fan et al. (1) describes the generation of multivalent antibody fragments using collagen sequence-based peptides to drive multimerization. In 2006, our group reported in The International Journal of Cancer that fusion of the Nterminal association subdomain (50 residues) of murine collagen XVIII NC1, responsible for noncovalent trimerization of collagen XVIII alpha chains, to the C-terminus of a single-chain variable fragment (scFv) confers their natural trimeric state to the fused antibody (2). The homo-trimeric molecules were isolated in functional active form from the cell culture supernatant of gene-modified human cells and showed high stability in the presence of relevant proteinases (2). The scFv used recognizes an angiogenesis-associated laminin epitope (3) and inhibits tumor angiogenesis and growth in vivo (4). We demonstrated that trimeric scFv-NC1 bound to laminin, and was more effective in blocking capillary morphogenesis in vitro, and in preventing tumor growth in vivo than its monomeric version (2). Given the wide availability of Internet-based search engines, we regret the omission in the above mentioned paper of our original reference showing for the first time the ability of collagen-derived sequences to promote antibody trimerization.

Diet &amp; Nutrition: Acrylamide Study Suggests Breast Cancer Link

The International Agency for Research on Cancer classifies acrylamide as a probable human carcinogen. It has only been in recent months that an epidemiologic study first found a link between dietary acrylamide and human cancer risk. Now Danish researchers report that acrylamide adduct levels in blood are associated with an increased risk of breast cancer in postmenopausal women. This is the first epidemiologic study to use blood biomarkers to assess acrylamide exposure. The findings, says first author Pelle Thonning Olesen, emphasize the importance of using biomarkers for exposure assessment. “Biomarkers are a more trustworthy indicator for exposure,” he says. Before 2002, people were known to be exposed to acrylamide in certain industries and through smoking tobacco. That year, the Swedish National Food Administration discovered that acrylamide also forms in fried or baked starchy foods such as french fries, coffee, and baked goods. Diet is now thought to be the major source of exposure among nonsmokers, but the cancer risk posed by acrylamide in food is unknown. All previous epidemiologic trials estimated acrylamide consumption from food frequency questionnaires. Olesen, a toxicologist at the Technical University of Denmark, Soborg, and colleagues instead measured levels of acrylamide and a key metabolite, glycidamide, bound to hemoglobin. The subjects included 374 postmenopausal women with breast cancer and 374 controls who participated in the Danish Cancer Society’s Diet, Cancer, and Health Study. Adduct levels of acrylamide among smokers reflect both dietary and smoking intake of the compound. After statistical adjustments for smoking behavior, women with the highest acrylamide-hemoglobin levels showed a 2.7 times higher risk of estrogen receptor–positive breast cancer compared with women with the lowest acrylamide-hemoglobin levels. The risk rose with increasing acrylamide exposure. Acrylamide-hemoglobin levels were not linked to estrogen receptor–negative breast cancer, and glycidamide-hemoglobin levels showed no connection with any breast cancer. The finding that only acrylamide-hemoglobin was associated with breast cancer suggests that the compound may induce cancer by nongenotoxic routes such as alkylation of proteins that could alter estrogen receptor function. The findings were reported in the 1 May 2008 issue of the International Journal of Cancer. “This is an important study because it’s the first to measure acrylamide adducts,” says epidemiologist Lorelei Mucci, an assistant professor at the Harvard School of Public Health. Nonetheless, the study should be repeated in larger numbers of nonsmoking women, according to Mucci, because more than half the cases and controls were current or former smokers. “The big public health question here is whether the amount of acrylamide in foods is enough to lead to cancer,” Mucci says. It is possible that other chemical compounds formed along with acrylamide may be the culprit in any cancer link. “Acrylamide-hemoglobin may be a biomarker for other carcinogenic chemicals formed during the heating of foods,” cautions Olesen.

Where we publish

Pathologists provide the link between the clinical and the natural science aspect of medicine and are often integrated in translational research projects. We wanted to analyse the spread and position of research led by pathologists in the scientific literature. Publications from institutes of pathology of 21 randomly chosen German university cities between 2004 and 2006 were analysed. To validate our findings we subsequently extended our analysis to 45 university cities from Italy, France and the Nordic countries (Denmark, Finland, Iceland, Norway and Sweden). We detected publications directed by pathologists in numerous biomedical journals, most of which not categorised as "pathology" journals. In the analyses from Germany and Italy, Virchows Archiv led the ranking in terms of absolute number of publications. Meanwhile, Anticancer Research and the International Journal of Cancer took the lead concerning publications from pathologists from the Nordic countries. Our results mirror the wide diversification in scientific pathology and the overlap with other medical subspecialties, especially oncology, cell biology and biochemistry. This explains the problem of defining "pathology" journals and deducting pathological research activity from these publications. However, the general notion that pathology journals are the most important communication medium of pathology research was affirmed.

The role of microRNAs in human cancer

Non-coding RNAs have recently been identified as important regulatory elements in mammalian and non-mammalian cells. In human cells, the number of microRNAs has been expected to exceed the one thousand mark. In cancer research, the interest in this class of small RNA molecules is particularly pronounced. This originates from aberrant expression patterns of specific microRNAs in various forms of human cancers as well as from proposals that specific microRNAs contribute to oncogenesis by either fulfilling tumorsuppressor functions or acting themselves as oncogenic factors. The expression profiles of microRNAs seem to vary in different types of presently analyzed human cancers, although characteristic patterns have been established for specific types of carcinomas. Besides their role in basic mechanisms leading to oncogenesis, they already start to play an increasing role in the diagnosis, prognosis and response to treatment of cancer patients. It is somewhat likely that their role will be extended in the future to therapeutic approaches, in particular to the identification of new therapeutic targets. The International Journal of Cancer wishes to emphasize the importance of this rapidly evolving field . The “Special Section” of this issue presents two contributions from pioneers in this field, George Calin from the MD Anderson Cancer Center in Houston, Texas, and Lin Zhang, Department of Obstetrics, University of Pennsylvania, Philadelphia, and their respective co-workers. The report by the Calin group emphasizes the role of miRNA profiling as diagnostic tools, whereas Zheng and colleagues stress the characterization of epigenetic regulation of microRNA as potential cancer biomarkers and as new therapeutic targets for future development. Jointly with my co-organizer of this Special Section, Webster Cavenee, I wish to express our gratitude to both groups for their contributions and trust that the International Journal of Cancer will provide a forum for future contributions in this exciting area of cancer research.

Research misconduct: learning the lessons

”The bulk of Jon Sudbø's scientific publications are invalid due to the fabrication and manipulation of the underlying data material.”1 This harsh conclusion sums up the investigation of a Norwegian researcher's complete scientific production. The investigation was started after a paper in The Lancet was shown to be based entirely on fabricated raw data.2,3 Several papers by the same author, published in journals such as The New England Journal of Medicine and International Journal of Cancer, were found fraudulent by the Investigation Commission.

Notice of Retraction: “Autonomous histopathological regression of primary tumours associated with specific immune responses to cancer antigens” (J Pathol 2003; 200: 383–395)

AbstractThe article to which this notice applies [DOI: 10.1002/path.1369], by Saleh FH, Crotty KA, Hersey P, Menzies SW and Rahman W has been retracted by agreement between the authors, the journal Editor‐in‐Chief, C Simon Herrington, and John Wiley &amp; Sons, Ltd. The retraction has been agreed due to substantial overlap between this article and the following article published in the International Journal of Cancer “Primary melanoma tumour regression associated with an immune response to the tumour‐associated antigen melan‐A/MART‐1”, by Saleh FH, Crotty KA, Hersey P and Menzies SW; Volume 94; 2001, pages 551–557.

Viewing a Stressful Episode of ER

See related article, pages 1186–1193 How cardiac cells sense, respond, and adapt to acute and chronic changes of their metabolic and environmental milieu remain among the most enigmatic and fundamental questions in contemporary cardiovascular biology and medicine. With more than 700 000 deaths and 6 million hospital discharges annually in the U.S., ischemic heart disease remains a major public health problem. If prompt therapy is delayed, myocardial injury from depletion of high-energy phosphates from inhibition of oxidative phosphorylation is inevitable. Ischemia/reperfusion (I/R) also unleashes a cascade of cellular and molecular events whose sequalae may sustain organ function at diminished capacity but, beyond a critical balance, threatens the organism’s survival. Reactive oxygen and nitrogen species (ROS; RNS), released from the mitochondria and other sources, alter the tertiary and quaternary structures of proteins, exposing their hydrophobic residues to allow conformational tendencies toward protein misfolding and aggregation.1 Nevertheless, highly sophisticated schemes equipped with molecular sensors, rapid response pathways, and adaptor mechanisms have evolved to mitigate the accumulation of unfolded proteins in a compartment specific manner, principally the cytoplasm and endoplasmic reticulum (ER). Although the cardiovascular field has paid considerably more attention to the cytoprotective mechanisms of heat shock proteins in response to I/R, termed the “classical” heat shock response, a parallel series of events has been unfolding at a breathtaking pace in the ER, with arguably considerable excitement to warrant a sneak preview of the article published in this issue of Circulation Research . To place into perspective the current buzz being garnered by the ER requires a brief discussion of more familiar themes in the field, which were developed thanks to the pioneering efforts of cardiovascular scientists, and others, working on the complementary stress response network, best exemplified by heat shock “stress” proteins and their transcriptional regulators. First discovered by …