• Survival improvement in thoracic cancer: progress from the last decade and beyond.

      Manegold, C; Thatcher, Nick; Heidelberg University, Interdisciplinary Thoracic Oncology and Department of Surgery, University Medical Center Mannheim, Theodor-Kutzer-Ufer 1-3, 68167 Mannheim, Germany. christian.manegold@chir.ma.uni-heidelberg.de (2007-08)
      Lung cancer remains a major concern, being the most common cancer worldwide, and with the incidence continuing to increase in many countries. Survival following diagnosis of non-small cell lung cancer (NSCLC) remains relatively low, with 1-year overall survival being approximately 25%. However, the development of new drugs and innovative treatment approaches is improving the outlook for patients with NSCLC. Important contributions have been made to the treatment of NSCLC and continue to do so with the development of gemcitabine, and more recently, pemetrexed. Major advances in the understanding of cancer biology have identified many potential targets for rationally designed novel therapies. Many of these targets are components of signalling pathways or metabolic processes that contribute to one of the hallmarks of the cancer phenotype. The novel targeted therapy enzastaurin, which is a derivative of the prototypical protein kinase C inhibitor staurosporine, is currently being investigated as monotherapy in NSCLC following first- or second-line chemotherapy. Research into the use of pharmacogenomics to tailor therapy to the individual patient is another innovative field that is yielding promising results that may help to improve the clinical outcome for patients with NSCLC in the future.
    • Survival in patients with multiple primary melanomas: Systematic review and meta-analysis

      Peek, G.; Olsen, C. M.; Baade, P.; Youlden, D. R.; Aitken, J. F.; Green, Adèle C; Khosrotehrani, K.; Queensland Skin and Cancer Foundation, Queensland Institute of Dermatology, South-Brisbane, Queensland, Australia. (2020)
      Background: The literature surrounding survival of patients with multiple primary melanomas (MPM) yields variable and opposing findings, constrained by statistical challenges. Objectives: To critically examine the available literature regarding survival of patients with MPM compared with a single primary melanoma and detail statistical methods used. Methods: Electronic searches were performed of PubMed, Embase, Web of Science, and Scopus, with cross-checking of references, for the period January 1956 to June 2019. Studies published in English examining survival in patients with multiple melanomas were included. Case studies and small case series were excluded. Results: There were 14 studies eligible for inclusion. Conclusions on survival varied markedly depending on the statistical method used. Four studies that accounted for survival bias by partitioning the survival time were included in the quantitative review, with 3 of these reporting a survival disadvantage for MPM, whereas the fourth showed no difference in survival. The pooled hazard ratio was 1.39 (95% confidence interval, 1.07-1.81) but with significant heterogeneity (I2 = 96.8%, Phet < .001). Limitations: Studies showed significant heterogeneity in methodology. Conclusion: When data were analyzed with robust statistical methods, patients with MPM had a survival disadvantage compared with patients with a single primary melanoma.
    • Survival of cells in mammalian tissues after low doses of irradiation: a short review.

      Hendry, Jolyon H; Department of Radiobiology, Paterson Institute for Cancer Research, Christie Hospital, Manchester, U.K. (1988-01)
      A survey of data in the literature indicates that the radiosensitivity of cells to doses less than 1 Gy varies widely within cell lineages and less so between lineages. This is due in large part to the differentiation status and division capacities of the cells, and possibly also to the grouping of cells into 'viable units'. In addition, the mode of cell death is important, and cells susceptible to natural apoptosis are particularly radiosensitive. There are also quite marked differences in cell sensitivity between species.
    • The survival of differentiating embryonic stem cells is dependent on the SCF-KIT pathway.

      Bashamboo, Anu; Taylor, A Helen; Samuel, Kay; Panthier, Jean-Jacque; Whetton, Anthony D; Forrester, Lesley M; John Hughes Bennett Laboratory, Edinburgh Cancer Centre, University of Edinburgh, Western General Hospital, Crewe Road, Edinburgh, EH4 2XU, UK. (2006-08-01)
      The stem cell factor (SCF)-KIT signal transduction pathway plays a role in the proliferation, differentiation and survival of a range of stem and progenitor cell types but little is known about its function in embryonic stem (ES) cells. We generated ES cells carrying a null allele of Kit as well as a knock-in allele that encodes an SCF-independent hybrid KIT receptor that can be activated by the FKBP binding drug, AP20187. KIT null ES cells die when induced to differentiate upon withdrawal of leukaemia inhibitory factor in monolayer culture. This phenotype is recapitulated in wild-type ES cells treated with a KIT-neutralising antibody and reversed in mutant cells by activation of the hybrid KIT receptor. Differentiating KIT null ES cells exhibit elevated levels of DNA laddering and reduced BCL2 expression, indicative of apoptosis. We conclude that mouse ES cell differentiation in vitro is dependent on the SCF-KIT pathway contrasting with the apparently normal differentiation of KIT null inner cell mass or epiblast cells in vivo. This discrepancy could be explained by the presence of compensatory signals in the embryo or it could lend support to the idea of a phenotypic relationship between ES cells and early germ cells.
    • Survival of head and neck cancer cells relies upon LZK Kinase-mediated stabilization of mutant p53

      Edwards, Zoe C; Trotter, Eleanor W; Torres-Ayuso, Pedro; Chapman, Phil; Wood, H; Nyswaner, Katherine; Brognard, John; Signalling Networks in Cancer, Cancer Research UK Manchester Institute (2017-07-31)
      Head and neck squamous cell carcinoma (HNSCC) includes epithelial cancers of the oral and nasal cavity, larynx, and pharynx and accounts for ∼350,000 deaths per year worldwide. Smoking-related HNSCC is associated with few targetable mutations but is defined by frequent copy-number alteration, the most common of which is gain at 3q. Critical 3q target genes have not been conclusively determined for HNSCC. Here, we present data indicating that MAP3K13 (encoding LZK) is an amplified driver gene in HNSCC. Copy-number gain at 3q resulted in increased MAP3K13 mRNA in HNSCC tumor samples and cell lines. Silencing LZK reduced cell viability and proliferation of HNSCC cells with 3q gain but not control cell lines. Inducible silencing of LZK caused near-complete loss of colony-forming ability in cells harboring 3q gain. These results were validated in vivo by evidence that LZK silencing was sufficient to reduce tumor growth in a xenograft model of HNSCC. Our results establish LZK as critical for maintaining expression of mutant stabilized p53. Cancer Res; 1-12. ©2017 AACR.
    • Survival of intestinal crypts after treatment by adriamycin alone or with radiation.

      Moore, James V; Broadbent, D A; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester (1980-11)
      A survival curve has been established for jejunal crypts of BDF1 mice treated by single i.p. doses of the antibiotic agent adriamycin. The threshold dose and Do were twice that for marrow CFU-S of these mice. The overall extrapolation number of the crypt survival curve was very low (1.3 +/- 0.13) compared to the value for gamma radiation. This observation is discussed with respect to the interpretation of crypt survival curves. We were unable to demonstrate any enhancement by adriamycin (reduction in Dq) of the response of microcolony-forming cells to radiation given immediately before the drug.
    • Survival of patients with early invasive melanoma down-staged under the new eighth edition of the American Joint Committee on Cancer staging system.

      von Schuckmann, L; Hughes, M; Lee, R; Lorigan, Paul C; Khosrotehrani, K; Smithers, M; Green, Adèle C; Population Health Department, Queensland Institute of Medical Research Berghofer Medical Research Institute, Herston, Australia (2019)
    • Surviving chromosome replication: the many roles of the S-phase checkpoint pathway.

      Labib, Karim; De Piccoli, Giacomo; Paterson Institute for Cancer Research, University of Manchester, Wilmslow Road, Manchester M20 4BX, UK. klabib@picr.man.ac.uk (2011-12-27)
      Checkpoints were originally identified as signalling pathways that delay mitosis in response to DNA damage or defects in chromosome replication, allowing time for DNA repair to occur. The ATR (ataxia- and rad-related) and ATM (ataxia-mutated) protein kinases are recruited to defective replication forks or to sites of DNA damage, and are thought to initiate the DNA damage response in all eukaryotes. In addition to delaying cell cycle progression, however, the S-phase checkpoint pathway also controls chromosome replication and DNA repair pathways in a highly complex fashion, in order to preserve genome integrity. Much of our understanding of this regulation has come from studies of yeasts, in which the best-characterized targets are the stimulation of ribonucleotide reductase activity by multiple mechanisms, and the inhibition of new initiation events at later origins of DNA replication. In addition, however, the S-phase checkpoint also plays a more enigmatic and apparently critical role in preserving the functional integrity of defective replication forks, by mechanisms that are still understood poorly. This review considers some of the key experiments that have led to our current understanding of this highly complex pathway.
    • Susceptibility of human leukaemias to cell-mediated cytotoxicity by interferon-treated allogeneic lymphocytes.

      Moore, Michael; Taylor, G; White, W J; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester M20 9BX, UK (1982)
      Twenty-two human leukaemias, comprising acute phase leucocytes from 13 acute myeloid and nine lymphoid leukaemias, were tested for susceptibility to spontaneous cell-mediated cytotoxicity (CMC) by untreated lymphocytes and lymphocytes treated for 18 h with 250 IU lymphoblastoid (Namalva) interferon (IFN-alpha). IFN-amplified killing (IAK) by lymphocytes from 24 normal lymphocyte donors was checked on the K562 erythroleukaemia cell line, for comparison with IAK on fresh leukaemias. Nine leukaemias were tested with lymphocytes from three donors, nine with lymphocytes from six donors, three with lymphocytes from nine donors, and one with lymphocytes from 11 donors. Some degree of susceptibility to IAK was found in five acute myeloid and five lymphoid leukaemias, which was markedly dependent upon the source of the effector lymphocytes and did not correlate with the degree of IAK on K562. The 12 other leukaemias were virtually resistant to IAK. The results emphasize the variability in the capacity of IFN-treated lymphocytes to lyse leukaemias that have not been adapted to tissue culture. The basis of effector recognition of cell line and fresh tumour targets is discussed.
    • Susceptibility of skin fibroblasts from individuals genetically predisposed to cancer, to transformation by the tumour promoter 12-O-tetradecanoylphorbol-13-acetate.

      Gainer, H S; Schor, Seth L; Kinsella, Anne R; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Wilmslow Road, Withington, Manchester M20 9BX, UK (1984-09-15)
      Skin fibroblasts from patients with hereditary retinoblastoma (RB) and familial polyposis coli (FPC) were chosen for study since their predisposition to the tumour may be due to an inherited "initiation" event which is present in every cell. Thus, it might be predicted that skin fibroblasts from these patients would exhibit increased susceptibility to in vitro transformation by tumour promoters alone. In the case of skin fibroblasts from RB patients, transformation as assessed by the ability of the cells to grow in semi-solid medium and their migration in collagen gels did not occur. However, experiments involving skin fibroblasts from FPC patients showed certain of these cells to grow in semi-solid medium following treatment with the tumour promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) alone, although the pattern of migration of the parent cell population in collagen gels was unchanged and they were non-tumorigenic in nude mice. The clones which grew in semi-solid medium, although stable with regard to anchor-age-independent growth, were also unaltered in terms of their migration pattern in collagen gels and their tumorigenicity in nude mice, and were considered not to be completely transformed. Parallel cytogenetic analysis showed that, during the course of these transformation studies, TPA significantly increased not only tetraploidy but also the chromosome aberration frequency. Several quadriradial figures were noted.
    • Sustained tumour eradication after induced caspase-3 activation and synchronous tumour apoptosis requires an intact host immune response.

      Melis, M H M; Simpson, Kathryn L; Dovedi, Simon J; Welman, Arkadiusz; MacFarlane, M; Dive, Caroline; Honeychurch, Jamie; Illidge, Timothy M; Targeted Therapy, Institute of Cancer Sciences, Manchester Academic Health Science Centre, University of Manchester, Manchester, UK. (2013-05)
      Effective anticancer treatments often result in the induction of large amounts of tumour cell death. In vivo, such dying tumour cells are a potential source of antigens for T-cell stimulation. Although apoptosis is generally considered nonimmunogenic, recent evidence suggests that some anticancer therapies that induce apoptosis can elicit antitumour immune responses. Here, a doxycycline-inducible, constitutively active caspase-3 ('death switch') was constructed in a murine tumour model to explore the impact of the host immune response to rapid, synchronous and substantial tumour cell apoptosis. In vitro, up to 80% of tumour cells underwent apoptotic cell death within 24 h and death was accompanied by the release of potential 'danger signal' molecules HMGB1 and HSP90. In vivo, death switch induction provoked rapid, pronounced tumour regression in immune-competent and immune-deficient mice, but sustained tumour eradication was observed only in immune-competent mice. Moreover, the majority of mice that were tumour free after death switch induction were protected from further tumour rechallenge. In addition, long-term remission after induction of the death switch was completely abrogated following depletion of CD8 T cells. These data suggest that sustained tumour eradication after substantial tumour apoptosis requires an antitumour host immune response that prevents tumour relapse. In many patients, cancer therapies produce encouraging initial responses that are only short lived. These results provide new insights that may have important implications for further development of strategies that result in long-term tumour clearance after initially effective anticancer treatment.
    • SYBR Green I staining of pulsed field agarose gels is a sensitive and inexpensive way of quantitating DNA double-strand breaks in mammalian cells.

      Kiltie, Anne E; Ryan, Anderson J; Department of Experimental Radiation Oncology, Paterson Institute for Cancer Research, Manchester M20 4BX, UK. (1997-07-15)
      Pulsed field gel electrophoresis (PFGE) is widely used to measure DNA double strand breaks (dsb). The DNA of cultured cells can be prelabelled with radioactivity, which helps greatly in detection and quantitation of DNA dsb. However, this approach cannot be used with non-cycling cells from biopsy material. We describe a method which uses SYBR Green I to stain DNA in dried agarose gels. DNA is detected and analysed using readily available camera equipment and image analysis software. This method is as sensitive as [3H]thymidine prelabelling of cells and allows DNA dsb to be measured simply and economically in non-cycling cells.
    • Synchronizing Progression of Schizosaccharomyces pombe Cells from G2 through Repeated Rounds of Mitosis and S Phase with cdc25-22 Arrest Release.

      Hagan, Iain M; Grallert, Agnes; Simanis, V; CRUK Cell Division Group, Cancer Research UK Manchester Institute, University of Manchester, Manchester M20 4BX, (2016)
      Transient inactivation of the cdc25(+) gene product by manipulation of the culture temperature for cdc25-22 cells is the most commonly exploited approach to mitotic synchronization in fission yeast. Because Cdc25 removes the inhibitory phosphate placed on Cdk1 by Wee1, inactivation of Cdc25 arrests cells at the G2/M boundary. Incubation at the restrictive temperature of 36°C for just over one generation time forces all cells in the culture to accumulate at the G2/M boundary. Restoration of Cdc25 function via a return to the permissive temperature or chemical inhibition of Wee1 activity at 36°C can then promote a highly synchronous wave of cell division throughout the culture. These approaches can be performed on any scale and thus support simultaneous assessment of numerous events within a single culture. After describing this simple and widely applicable procedure, we discuss frequently overlooked issues that can have a considerable impact on the interpretation of data from cdc25-22 induction-synchronized cultures.
    • Synchronizing Progression of Schizosaccharomyces pombe Cells from Prophase through Mitosis and into S Phase with nda3-KM311 Arrest Release.

      Hagan, Iain M; Grallert, Agnes; Simanis, V; CRUK Cell Division Group, Cancer Research UK Manchester Institute, University of Manchester, Manchester M20 4BX, (2016)
      Here, we describe how the rapid reversibility of the nda3-KM311 cold-sensitive β-tubulin mutation was optimized by Mitsuhiro Yanagida's laboratory to synchronize mitotic progression in an entire cell population. The inability to form microtubules following the loss of β-tubulin function at 20°C triggers the spindle assembly checkpoint, which arrests mitotic progression. Restoration of β-tubulin function by rewarming to 30°C (or higher) releases the arrest, generating a highly synchronous progression through mitosis. The viability of nda3-KM311 strains at 30°C makes it feasible to generate double mutants between nda3-KM311 and any temperature-sensitive mutant that can also grow at 30°C. These double mutants can be used in reciprocal shift analyses, in which cold-induced early mitotic arrest is relieved by a shift to 36°C, which then inactivates the product of the second mutant gene. The addition of microtubule depolymerizing drugs before the return to 36°C will maintain checkpoint signaling at 36°C transiently, permitting analysis of the impact of temperature-sensitive mutations on checkpoint function. Silencing the checkpoint of nda3-KM311-arrested cells at 20°C through chemical inhibition of aurora kinase is a powerful way to study checkpoint recovery pathways and mitotic exit without anaphase.
    • Syndecan-1 and -4 synthesized simultaneously by mouse mammary gland epithelial cells bear heparan sulfate chains that are apparently structurally indistinguishable.

      Zako, Masahiro; Dong, Jianying; Goldberger, Olga; Bernfield, Merton; Gallagher, John T; Deakin, Jon A; Division of Newborn Medicine, Children's Hospital, Harvard Medical School, Boston, Massachusetts 02215, USA. zako@aichi-med.ac.jp (2003-04-11)
      Many of the biological functions attributed to cell surface heparan sulfate (HS) proteoglycans, including the Syndecan family, are elicited through the interaction of their HS chains with soluble extracellular molecules. Tightly controlled, cell-specific sulfation and epimerization of HS precursors endows these chains with highly sulfated, iduronate-rich regions, which are major determinants of cytokine and matrix-protein binding and which are interspersed by N-acetylated, poorly sulfated regions. Until this study, there have been no comprehensive structural comparisons made on HS chains decorating simultaneously expressed, but different, syndecan core proteins. In this paper we demonstrate that the HS chains on affinity-purified syndecan-1 and -4 from murine mammary gland cells are essentially identical by a number of parameters. Size determination, disaccharide analyses, enzymatic and chemical scission methods, and affinity co-electrophoresis all failed to reveal any significant differences in fine structure, domain organization, or ligand-binding properties of these HS species. These findings lead us to suggest that the imposition of the fine structure onto HS occurs independently of the core protein to which it is attached and that these core proteins, in addition to the HS chains, may play a pivotal role in the various biological functions ascribed to these macromolecules.
    • Synergistic effects of imatinib (STI 571) in combination with chemotherapeutic drugs in head and neck cancer.

      Bruce, Iain A; Slevin, Nicholas J; Homer, Jarrod J; McGown, Alan T; Ward, Timothy H; Paterson Institute for Cancer Research, Christie Hospital, Manchester, UK. (2005-08)
      The tyrosine kinase inhibitor imatinib (STI 571; glivec) is a potent inhibitor of bcr-abl, c-kit and platelet-derived growth factor receptors. Imatinib was evaluated both alone and in combination with established chemotherapeutic agents in adenoid cystic carcinoma (ACC) primary cultures and established cell lines representing squamous cell carcinoma of the head and neck (HNSCC). Over 90% of ACC tumors are c-kit-positive, and these primary cultures proved to be of short-term usefulness in assessing chemosensitivity. Interaction was determined over a wide range of drug combinations using a statistical three-dimensional analysis model. Both ACC short-term cultures and HNSCC cell lines were demonstrated to have a response ranging from additive to synergistic when imatinib and cisplatin were combined. The interaction of imatinib on cisplatin-induced DNA cross-linking was further investigated using the comet-X assay. In contrast, significant antagonism was observed when imatinib and gemcitabine were combined. Since gemcitabine is activated by deoxycytidine kinase (dCK), the effect of imatinib on this enzyme was investigated. A dose-dependent inhibition of dCK was observed, highlighting this kinase as a possible additional secondary molecular target for imatinib. This work demonstrates a synergistic interaction between cisplatin and imatinib, which may prove to be clinically relevant in the future management of both ACC and HNSCC.
    • Synergistic interactions in haemopoiesis: biological implications and clinical use.

      Dexter, T Michael; Christie Hospital, Withington, Manchester, UK. (1993)
      Growth factors promote the survival and proliferation of haemopoietic stem and progenitor cells, and in their absence the haemopoietic cells undergo apoptosis and die. The results of studies reported here indicate that multipotent stem cells have receptors for most, if not all, of the growth factors, but that even saturated binding of the receptors for a single growth factor is not sufficient to transduce an effective stimulus for the proliferation of these cells (possibly due to very low receptor numbers). However, when the growth factors are combined synergistic effects can be seen. Studies in which stem cell factor was used in combination with other growth factors showed that stem cell factor allowed the survival of stem cells, while a second growth factor (granulocyte-macrophage colony-stimulating factor) stimulated the stem cells to develop normally. Stem cell factor was also shown to alter the dose-response relationships of developing haemopoietic cells for other growth factors.
    • A syngeneic mouse B-cell lymphoma model for pre-clinical evaluation of CD19 CAR T cells.

      Kueberuwa, Gray; Zheng, W; Kalaitsidou, Milena; Gilham, David E; Hawkins, Robert E; Manchester Cancer Research Centre Building, Department Cancer Sciences, University of Manchester (2018)
      The astonishing clinical success of CD19 chimeric antigen receptor (CAR) T-cell therapy has led to the approval of two second generation chimeric antigen receptors (CARs) for acute lymphoblastic leukemia (ALL) andnon-Hodgkin lymphoma (NHL). The focus of the field is now on emulating these successes in other hematological malignancies where less impressive complete response rates are observed. Further engineering of CAR T cells or co-administration of other treatment modalities may successfully overcome obstacles to successful therapy in other cancer settings. We therefore present a model in which others can conduct pre-clinical testing of CD19 CAR T cells. Results in this well tested B-cell lymphoma model are likely to be informative CAR T-cell therapy in general. This protocol allows the reproducible production of mouse CAR T cells through calcium phosphate transfection of Plat-E producer cells with MP71 retroviral constructs and pCL-Eco packaging plasmid followed by collection of secreted retroviral particles and transduction using recombinant human fibronectin fragment and centrifugation. Validation of retroviral transduction, and confirmation of the ability of CAR T cells to kill target lymphoma cells ex vivo, through the use of flow cytometry, luminometry and enzyme-linked immunosorbent assay (ELISA), is also described. Protocols for testing CAR T cells in vivo in lymphoreplete and lymphodepleted syngeneic mice, bearing established, systemic lymphoma are described. Anti-cancer activity is monitored by in vivo bioluminescence and disease progression. We show typical results of eradication of established B-cell lymphoma when utilizing 1st or 2nd generation CARs in combination with lymphodepleting pre-conditioning and a minority of mice achieving long term remissions when utilizing CAR T cells expressing IL-12 in lymphoreplete mice. These protocols can be used to evaluate CD19 CAR T cells with different additional modification, combinations of CAR T cells and other therapeutic agents or adapted for the use of CAR T cells against different target antigens.
    • The syntheses and properties of tricyclic pyrrolo[2,3-d]pyrimidine analogues of S6-methylthioguanine and O6-methylguanine

      Hornillo-Araujo, Ana R; Burrell, Adam J M; Aiertza, Miren K; Shibata, Takayuki; Hammond, David M; Edmont, Dolores; Adams, Harry; Margison, Geoffrey P; Williams, David M; Centre for Chemical Biology, Richard Roberts Building, Department of Chemistry, University of Sheffield, Brook Hill, Sheffield, UK S3 7HF (2006)
    • Synthesis and anticancer activities of 4-oxobenzopyrano[2,3-d]pyrimidines.

      Hadfield, John A; Pavlidis, V H; Perry, P J; McGown, Alan T; Cancer Research Campaign Section of Drug Development and Imaging, Paterson Institute for Cancer Research, Manchester, UK. jhadfield@picr.man.ac.uk (1999-07)
      Several 2-aryl-4-oxoxbenzopyrano[2,3-d]pyrimidines have previously been shown to exhibit in vivo antitumor activity in mice with P388 lymphocytic leukemia. In the present study, a series of novel substituted benzopyrano[2,3-d]pyrimidines have been prepared and tested for cytotoxic activity against a panel of cancer cell lines including the P388 lymphocytic leukemia cell line. The unsubstituted parent compound, some methoxylated derivatives and a cyclohexyl derivative all exhibited potent cytotoxic activity (IC50 values 0.3-0.64 microM). A number of derivatives, including the unsubstituted parent pyrimidine, were shown to cause a significant perturbation in cell cycle kinetics with an observed 2- to 3-fold increase in cells in the G2/M phase of the cell cycle. Furthermore, a polymethoxylated derivative, 2-(3,4,5-trimethoxyphenyl)-9-methoxy-4-oxo-2,3-dihydrobenzopyrano[ 2,3-d]pyrimidine 13, was shown to be selectively active against a number of human ovarian cell lines.