• Cyclophosphamide as an adjuvant to X-rays in treatment of a radioresistant solid tumor of the rat, hepatoma H-4-II-E.

      Moore, James V; Rowley, Roy; Hopkins, Harold A; Ritenour, E Russell; Looney, William B; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester (1979-09)
    • Cyclophosphamide decreases O6-alkylguanine-DNA alkyltransferase activity in peripheral lymphocytes of patients undergoing bone marrow transplantation.

      Lee, Siow Ming; Crowther, Derek; Scarffe, J Howard; Dougal, Mark; Elder, Rhoderick H; Rafferty, Joseph A; Margison, Geoffrey P; CRC Department of Carcinogenesis, Paterson Institute for Cancer Research, Manchester, UK. (1992-08)
      O6-alkylguanine-DNA-alkyltransferase (ATase) levels were measured in extracts of peripheral blood lymphocytes taken at various times during chemotherapy from 19 patients with various haematological malignancies. Seven patients with advanced Hodgkin's disease received preparative treatment consisting of cyclophosphamide (1.5 g m-2, daily) administered on days 1 to 4 and BCNU (600 mg m-2) on day 5 prior to autologous bone marrow rescue (ABMR) delivered on day 7. Treatment in the remaining 12 patients consisted of cyclophosphamide (1.8 g m-2, daily) given on days 1 and 2 followed at day 4 with total body irradiation (TBI) administered in six fractions over the subsequent 3 days to a total dose of 1200 cGy prior to bone marrow transplantation. In the Hodgkin's group, significant decreases in ATase activity were seen during the cyclophosphamide treatment, and the median ATase nadir was 32% (range 0% to 57%) of pretreatment levels following 4 days of cyclophosphamide. In one patient, no ATase activity was detectable following the 4th cyclophosphamide treatment. ATase activities decreased further after BCNU administration to a median of 19% (range 0% to 32%) of pretreatment levels. Extensive cyclophosphamide-induced reduction of lymphocyte ATase levels was also seen in the other group of 12 patients treated with cyclophosphamide/TBI: postcyclophosphamide median ATase nadir was 35% (range 12% to 78%) of the pretreatment levels. No ATase depletion was seen when cyclophosphamide (up to 10 mM) was incubated for 2 h with pure recombinant human ATase in vitro whereas ATase activity was reduced by 90% on preincubation with 100 microns acrolein or with greater than 1 mM phosphoramide mustard. This suggests that a cyclophosphamide-induced decrease in ATase levels in human peripheral lymphocytes in vivo may be due to depletion mediated by the production of intracellular acrolein. Since ATase appears to be a principal mechanism in cellular resistance to the cytotoxic effects of BCNU and related alkylating agents, these observations suggest that a cyclophosphamide-induced reduction in ATase activity may be an additional factor in the effectiveness of the combined sequential therapy.
    • Cyclophosphamide inhibition of anti-CD40 monoclonal antibody-based therapy of B cell lymphoma is dependent on CD11b+ cells.

      Honeychurch, Jamie; Glennie, Martin J; Illidge, Timothy M; Cancer Research UK Oncology Unit, Tenovus Research Laboratory, Cancer Sciences Division, School of Medicine, Southampton General Hospital, Southampton, Hampshire. (2005-08-15)
      Monoclonal antibody (mAb)-based immunotherapy is now established as an important option for treating some cancers. The antitumor effects may be further enhanced by combining mAb with conventional chemotherapy. Certain novel immunomodulatory mAbs such as anti-CD40 have shown significant activity in preclinical models. We therefore assessed the efficacy of combining anti-CD40 mAb, known to elicit CTL responses against murine lymphoma models with the commonly used cytotoxic drug, cyclophosphamide. Using the syngeneic tumor model, BCL1, we have shown that timing of cyclophosphamide relative to mAb is critical to therapeutic outcome. Pretreatment with cyclophosphamide 7 to 10 days prior to mAb results in markedly reduced survival levels, similar to that achieved with cyclophosphamide alone. Conversely, when anti-CD40 is given before cyclophosphamide, the level of tumor protection was moderately increased. In vivo tracking experiments reveal that pretreatment with cyclophosphamide leads to diminished CTL expansion, as well as an increased number of CD11b+ cells that display an activated phenotype. These latter cells are able to inhibit T-cell proliferation, at least in part via production of nitric oxide, but do not induce T-cell apoptosis. Furthermore, adoptive transfer of the induced CD11b+ cells is sufficient to inhibit anti-CD40 therapy in tumor-bearing recipients. We have shown that the timing of cyclophosphamide relative to mAb administration is critical to the therapeutic outcome, and although the combination can improve survival, cyclophosphamide given prior to immunotherapy may generate a population of myeloid cells that can interfere with CTL responses and compromise the therapeutic outcome.
    • Cysteine cathepsin protease inhibition: an update on its diagnostic, prognostic and therapeutic potential in cancer

      Soond, SM; Kozhevnikova, MV; Townsend, Paul A; Zamyatnin, AA; Institute of Molecular Medicine, Sechenov First Moscow State Medical University, Trubetskaya str. 8-2, 119991 Moscow, Russia. (2019)
      In keeping with recent developments in basic research; the importance of the Cathepsins as targets in cancer therapy have taken on increasing importance and given rise to a number of key areas of interest in the clinical setting. In keeping with driving basic research in this area in a translational direction; recent findings have given rise to a number of exciting developments in the areas of cancer diagnosis; prognosis and therapeutic development. As a fast-moving area of research; the focus of this review brings together the latest findings and highlights the translational significance of these developments.
    • Cytogenetic changes during the early stages of liver carcinogenesis in Chinese hamster: an in vivo--in vitro comparison.

      Swindell, J A; Ockey, Charles H; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester UK (1983-09)
      Cytogenetic changes were investigated during the early stages of hepatic adenocarcinoma development in Chinese hamsters injected with a single dose of dimethylnitrosamine (DMN). An in vivo-in vitro comparison was made from 7 to 35 weeks after injection. A partial hepatectomy was used to stimulate mitosis for in vivo analysis, and the excised liver, grown to the primary culture stage, was used for chromosome analysis in vitro. Aneuploidy, tetraploidy, and chromosome aberrations increased significantly in the hepatic cells of DMN-treated animals in vivo, with no significant change over the 7- to 35-week period. No differences, however, could be detected between the primary cultures of control and DMN-treated animals because of an inherent tendency for all cultures to develop aneuploid stem lines at an early stage in culture. A preferential involvement of chromosome #6 in the single trisomic state was demonstrated in vitro and to a minor extent in vivo. The relevance of increased aneuploidy in early carcinogenesis and the differences between the in vivo and in vitro results are discussed.
    • Cytogenetic studies of workers exposed to styrene: a review.

      Scott, David; Department of Cancer Genetics, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, United Kingdom. (1993)
      In 17 of 50 cytogenetic studies (on chromosomal aberrations, micronucleus formation and sister chromatid exchange) in peripheral blood lymphocytes from a total of 667 workers in industries in which there is exposure to styrene, significant increases in the frequency of chromosomal damage have been reported when compared with unexposed controls. The positive or negative outcome of these studies is, however, unrelated to the extent of exposure to styrene. Furthermore, in the 17 investigations in which positive results were found, there is no convincing evidence of a positive dose-response relationship, in spite of the wide ranges of exposure and different methods of measurement. There are also serious discrepancies between findings on the chromosomal damaging effects of styrene in human lymphocytes in vitro and the types of damage reported in exposed workers. The findings are not consistent with the interpretation that styrene is responsible for the observed effects in workers. Several other chemicals identified in the environment of styrene workers have been reported to induce chromosomal damage in vitro and/or in vivo.
    • A cytogenetic study of male breast cancer.

      Mitchell, Erika L D; Cancer Genetics Department, Paterson Institute for Cancer Research, Christie Hospital, Manchester, England. (1990-07-01)
      In a direct preparation from a male breast carcinoma two populations of cells were present, one hypodiploid (range 25-34) and the other hypertriploid (range 56-84). Twenty-two marker chromosomes were recognized. One of these, dic(5:11)(p14:q23) was present in one or two copies in every cell and has not been reported in any other case of breast cancer. There was a consistent monosomy of chromosome 7 and, in the hypertriploid cells, a gain of one to three copies of chromosome 3. The breakpoint 11q23 is a rare, folate-sensitive fragile site but was not expressed in peripheral blood cell lymphocytes from the patient.
    • Cytokeratin expression in epithelial cells isolated from the crypt and villus regions of the rodent small intestine.

      Flint, Neil; Pemberton, P W; Lobley, R W; Evans, Gareth S; CRC Department of Epithelial Biology, Paterson Institute for Cancer Research, Christie Hospital (NHS) Trust, Manchester, UK. (1994-01)
      Many physiological and structural features of epithelium in the small intestine are regulated during their transit from the crypt base to the villus tip. This crypt-villus axis is an important model for the study of the regulation of cell proliferation and differentiation. We have investigated the expression of cytokeratins in purified epithelial cells from the proliferative (crypt) and differentiated (villus) regions of this tissue. Three polypeptides were identified (cytokeratins 8, 18 and 19) as well as a fourth, 46 kDa polypeptide with similar electrophoretic characteristics to the recently identified cytokeratin 20. The distribution of these molecules was found to vary along the crypt-villus axis, with cytokeratin 18 being restricted to the proliferative crypt and cytokeratins 8 and 19 demonstrating more uniform distributions. The 46 kDa component was found to be expressed predominantly within the villus epithelium. Although there is no substantial evidence of a direct role for cytokeratins in the process of epithelial differentiation, these data suggest that differential expression of cytokeratins is associated with changes in intestinal epithelial differentiation.
    • Cytokine stimulation and the choice of promoter are critical factors for the efficient transduction of mouse T cells with HIV-1 vectors.

      Gilham, David E; Lie-A-Ling, Michael; Taylor, Naomi; Hawkins, Robert E; Cell Therapy Group, Cancer Research UK Department of Medical Oncology, University of Manchester, Manchester Academic Health Science Centre, Manchester, UK. dgilham@picr.man.ac.uk (2010-02)
      BACKGROUND: HIV-1 fails to successfully infect mouse T cells as a result of several blocks in the viral replication cycle. We investigated whether this also impacted on the use of HIV-1 derived lentiviral vectors for stable gene transfer into mouse T cells. METHODS: Freshly isolated primary mouse T cells were immediately mixed with lentiviral vectors encoding an enhanced green fluorescent protein marker gene and transduction frequency was determined after 5 days of culture. RESULTS: Optimal transduction required both mouse T cell activation and cytokine support. Furthermore, transduction was also dependent upon the promoter chosen, with the rank order of potency being PGK > EF1 > SFFV > CMV. HIV-1 lentiviral vectors also efficiently transduced cytokine-stimulated T cells (in the absence of antibody driven T cell activation), albeit with a lower level of transgene expression compared to fully-activated T cells. CONCLUSIONS: The present study demonstrates that primary mouse T cells can be efficiently transduced with HIV-1 lentiviral vectors, opening up prospects for their use in mouse models of gene-modified adoptive cellular therapy.
    • Cytokine-mediated protein kinase C activation is a signal for lineage determination in bipotential granulocyte macrophage colony-forming cells.

      Whetton, Anthony D; Heyworth, Clare M; Nicholls, S E; Evans, C A; Lord, J M; Dexter, T Michael; Owen-Lynch, P J; Department of Biochemistry and Applied Molecular Biology, UMIST, Manchester, United Kingdom. (1994-05)
      Granulocyte macrophage colony-forming cells (GM-CFC) have the potential to develop into either macrophages and/or neutrophils. With a highly enriched population of these cells we have found that although GM-CFC are equally responsive to macrophage colony stimulating factor (M-CSF) and stem cell factor (SCF) in terms of DNA synthesis, M-CSF stimulated the development of colonies containing macrophages in soft gel assays, while SCF promoted neutrophilic colony formation. When SCF and M-CSF were combined, mainly macrophage development was stimulated both in soft agar colony-forming assays and liquid cultures. An analysis of some potential signaling mechanisms associated with cytokine-mediated developmental decisions in GM-CFC revealed that M-CSF, but not SCF, was able to chronically stimulate phosphatidylcholine breakdown and diacylglycerol production, indicating that protein kinase C (PKC) may be involved in the action of M-CSF. Furthermore, M-CSF, but not SCF, can increase the levels of PKC alpha (PKC alpha) expression and stimulate the translocation of PKC alpha to the nucleus. When the PKC inhibitor, calphostin C, was added to GM-CFC cultured in M-CSF then predominantly neutrophils were produced, conversely PKC activators added with SCF stimulated macrophage development. The data indicate a role for PKC in M-CSF-stimulated macrophage development from GM-CFC.
    • The cytoplasmic filaments of the nuclear pore complex are dispensable for selective nuclear protein import.

      Walther, Tobias C; Pickersgill, Helen; Cordes, Volker C; Goldberg, Martin W; Allen, Terence D; Mattaj, Iain W; Fornerod, Maarten; Gene Expression Program, EMBL, D-69117 Heidelberg, Germany. (2002-07-08)
      The nuclear pore complex (NPC) mediates bidirectional macromolecular traffic between the nucleus and cytoplasm in eukaryotic cells. Eight filaments project from the NPC into the cytoplasm and are proposed to function in nuclear import. We investigated the localization and function of two nucleoporins on the cytoplasmic face of the NPC, CAN/Nup214 and RanBP2/Nup358. Consistent with previous data, RanBP2 was localized at the cytoplasmic filaments. In contrast, CAN was localized near the cytoplasmic coaxial ring. Unexpectedly, extensive blocking of RanBP2 with gold-conjugated antibodies failed to inhibit nuclear import. Therefore, RanBP2-deficient NPCs were generated by in vitro nuclear assembly in RanBP2-depleted Xenopus egg extracts. NPCs were formed that lacked cytoplasmic filaments, but that retained CAN. These nuclei efficiently imported nuclear localization sequence (NLS) or M9 substrates. NPCs lacking CAN retained RanBP2 and cytoplasmic filaments, and showed a minor NLS import defect. NPCs deficient in both CAN and RanBP2 displayed no cytoplasmic filaments and had a strikingly immature cytoplasmic appearance. However, they showed only a slight reduction in NLS-mediated import, no change in M9-mediated import, and were normal in growth and DNA replication. We conclude that RanBP2 is the major nucleoporin component of the cytoplasmic filaments of the NPC, and that these filaments do not have an essential role in importin alpha/beta- or transportin-dependent import.
    • Cytotoxic Michael-type amine adducts of alpha-methylene lactones alantolactone and isoalantolactone.

      Lawrence, Nicholas J; McGown, Alan T; Nduka, Jane; Hadfield, John A; Pritchard, Robin G; Department of Chemistry, UMIST, Manchester, UK. lawrencenj1@cardiff.ac.uk (2001-02-12)
      Two series of cytotoxic (IC50, K562 cell line, 1-24 microM) alpha-aminomethyl substituted lactones 3 and 4 were prepared by stereoselective Michael-type addition of amines to alantolactone (1) and isoalantolactone (2). The lactones 1 and 2 and their amine adducts induce apoptosis and act as alkylating agents.
    • Cytotoxicity of adherent cells associated with some human tumours and lung tissues.

      Vose, Brent M; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester (1978)
    • Cytotoxicity of the bioreductive agent RH1 and its lack of interaction with radiation.

      Kim, Joo-Young; West, Catharine M L; Valentine, Helen R; Ward, Timothy H; Patterson, Adam V; Stratford, Ian J; Roberts, Stephen A; Hendry, Jolyon H; Cancer Research UK Groups of Experimental Radiation Oncology, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, UK. (2004-03)
      BACKGROUND AND PURPOSE: RH1 is a new bioreductive agent that was developed as a cytotoxic agent with selectivity for tumour cells expressing high levels of the enzyme DT-diaphorase (DTD). The aim of the present study was to investigate the cytotoxicity of RH1 in relation to cellular levels of reducing enzymes and any interaction of RH1 with ionizing radiation under oxic and hypoxic conditions. PATIENTS AND METHODS: The MB-MDA231 human breast cancer cell line (WT) and WT cells transfected with the NQO1 gene encoding DTD (the D7 cell line) were used to examine the dependency of RH1's cytotoxicity on cellular DTD activity. The role of the 1-electron reducing enzyme P450 reductase was also studied using a P450 reductase-transfected isogenic cell line (R4). A clonogenic assay was used to investigate the cytotoxicity of RH1 with and without irradiation in air and in nitrogen. In all cases drug exposure was for 3 h. RESULTS: DTD levels were around 300-fold higher in D7 compared to WT and R4 cells. RH1 was cytotoxic at nanomolar concentrations to all the cell lines, and was 2-3 times more toxic in the D7 cells with high DTD than in the other two cell lines. Doses of RH1 was around 2-fold more effective in hypoxic than in oxic WT cells, but not by as much in D7 cells. RH1 did not radiosensitise the cells but showed an additive effect when combined with irradiation under oxic and hypoxic conditions. CONCLUSIONS: RH1 shows high clonogenic cytotoxicity to MDA231 cells with high DTD activity but its selectivity based on the presence of DTD is much less than as shown in previous reports. RH1 showed an additive cell killing effect when combined with irradiation under both oxic and hypoxic conditions.
    • D-Cycloserine destruction by alanine racemase and the limit of irreversible inhibition

      de Chiara C; Homsak M; Prosser, GA; Garza-Garcia A, Kelly G; Purkiss AG; Tate EW; de Carvalho LPS; Douglas, HL; Mycobacterial Metabolism and Antibiotic Research Laboratory, The Francis Crick Institute, London, UK. (2020)
    • Damage-induced apoptosis in intestinal epithelia from bcl-2-null and bax-null mice: investigations of the mechanistic determinants of epithelial apoptosis in vivo.

      Pritchard, D Mark; Potten, Christopher S; Korsmeyer, Stanley J; Roberts, Stephen A; Hickman, John A; CRC Molecular and Cellular Pharmacology Group, School of Biological Sciences, Stopford Building (G38), University of Manchester, Manchester M13 9PT, UK. (1999-12-02)
      The influence of bcl-2 and bax expression on apoptotic cell death in mouse intestinal epithelia was assessed using homozygously null mice. Apoptosis was induced in vivo by the enterotoxin 5-fluorouracil (5FU) or by gamma-irradiation and its cell positional incidence was assessed. 5FU and gamma-radiation treated bax-null mice surprisingly showed no reductions in apoptotic yield in the small intestine or midcolon at 4.5 h at cell positions in which both agents had previously been shown to strongly induce p53 protein expression. The colonic epithelia of 5FU treated bcl-2-null mice showed elevated levels of apoptosis at 4.5 h: from 48 apoptotic events in wild-type mice to 273 in the nulls, scoring 200 half crypts. The increase occurred specifically in the cell positions considered to harbour colonic stem cells, at the base of crypts, where there is selective expression of bcl-2. There was a modest but significant increase in apoptosis in the small intestine of the bcl-2-null mice although the epithelia of wild-type mice here are not immunohistochemically positive for bcl-2 protein. These findings show that bcl-2 plays a key role in determining the sensitivity of colonic stem cells to damage-induced death but that bax is not responsible for the p53-dependent induction of apoptosis in this context.
    • DCE-MRI biomarkers in the clinical evaluation of antiangiogenic and vascular disrupting agents.

      O'Connor, James P B; Jackson, Alan; Parker, Geoff J M; Jayson, Gordon C; Imaging Science and Biomedical Engineering, University of Manchester, Oxford Road, Manchester M13 9PT, UK.james.o'connor@manchester.ac.uk (2007-01-29)
      Dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) is now frequently used in early clinical trial assessment of antiangiogenic and vascular disrupting compounds. Evidence of drug efficacy and dose-dependent response has been demonstrated with some angiogenesis inhibitors. This review highlights the critical issues that influence T(1)-weighted DCE-MRI data acquisition and analysis, identifies important areas for future development and reviews the clinical trial findings to date.
    • Death of intestinal crypts and of their constituent cells after treatment by chemotherapeutic drugs.

      Moore, James V; Paterson Laboratories, CHristie Hospital and Holt Radium Institute, Manchester M20 9BX (1984-01)
      The number and spatial distribution of necrotic cells in the jejunal crypts of mice, has been measured after treatment by each of 6 cytotoxic drugs. At the LD10/8 day dose of each drug, the majority of necrotic cells were found below position 9 and numbers per crypt were similar for all drugs (approximately 8). These findings resemble those for radiation. However, major differences between agents were found in the calculated numbers of the microcolony-forming units (MFU) that determine overall crypt survival or ablation after high doses of cytotoxic agent. Numbers of MFU as assayed by radiation were approximately 80 per crypt, but only 2 when assayed by mechlorethamine hydrochloride, adriamycin and 5-fluorouracil, and 7 using BCNU. No crypts were destroyed by either cyclophosphamide or actinomycin D, despite the appearance of numerous necrotic cells in the lower part of the crypt. We conclude that in drug-treated intestine, necrotic cells may arise from a non-MFU compartment and the incidence and distributions of such cells are likely to be poor indicators of the response of the MFU.