• An extended Li-Fraumeni kindred with gastric carcinoma and a codon 175 mutation in TP53.

      Varley, Jennifer; McGown, Gail; Thorncroft, Mary R; Tricker, Karen J; Teare, M Dawn; Santibanez-Koref, Mauro F; Martin, J; Birch, Jillian M; Evans, D Gareth R; CRC Department of Cancer Genetics, Paterson Institute for Cancer Research, Manchester, UK. (1995-12)
      We present an extended family with Li-Fraumeni syndrome characterised by gastric and breast carcinoma, glioma, sarcoma, and leukaemia. This family showed strong evidence of linkage to TP53, and three of four tumours analysed showed loss of the wild type allele. A codon 175 missense mutation was identified in exon 5 in all available affected subjects. Counselling, screening, and issues surrounding presymptomatic testing are discussed.
    • Extended Middle East and North Africa: summary recommendations for the prevention of human papillomavirus infections and related cancers including cervical cancer.

      Seoud, M; Vaccarella, S; El-Kak, F; Sancho-Garnier, H; Jumaan, A; Kim, J; Garland, S; Stern, Peter L; LaMontagne, D; Albero, G; et al. (2013-12-30)
    • An extended set of PRDM1/BLIMP1 target genes links binding motif type to dynamic repression.

      Doody, Gina M; Care, Matthew A; Burgoyne, Nicholas J; Bradford, James R; Bota, Maria; Bonifer, Constanze; Westhead, David R; Tooze, Reuben M; Section of Experimental Haematology, Leeds Institute of Molecular Medicine, University of Leeds, Leeds LS9 7TF (2010-04-26)
      The transcriptional repressor B lymphocyte-induced maturation protein-1 (BLIMP1) regulates gene expression and cell fate. The DNA motif bound by BLIMP1 in vitro overlaps with that of interferon regulatory factors (IRFs), which respond to inflammatory/immune signals. At such sites, BLIMP1 and IRFs can antagonistically regulate promoter activity. In vitro motif selection predicts that only a subset of BLIMP1 or IRF sites is subject to antagonistic regulation, but the extent to which antagonism occurs is unknown, since an unbiased assessment of BLIMP1 occupancy in vivo is lacking. To address this, we identified an extended set of promoters occupied by BLIMP1. Motif discovery and enrichment analysis demonstrate that multiple motif variants are required to capture BLIMP1 binding specificity. These are differentially associated with CpG content, leading to the observation that BLIMP1 DNA-binding is methylation sensitive. In occupied promoters, only a subset of BLIMP1 motifs overlap with IRF motifs. Conversely, a distinct subset of IRF motifs is not enriched amongst occupied promoters. Genes linked to occupied promoters containing overlapping BLIMP1/IRF motifs (e.g. AIM2, SP110, BTN3A3) are shown to constitute a dynamic target set which is preferentially activated by BLIMP1 knock-down. These data confirm and extend the competitive model of BLIMP1 and IRF interaction.
    • Extensive amplification of single cells from CD34+ subpopulations in umbilical cord blood and identification of long-term culture-initiating cells present in two subsets.

      De Wynter, Erika A; Nadali, Gianpaolo; Coutinho, Lucia H; Testa, Nydia G; CRC Department of Experimental Haematology, Paterson Institute for Cancer Research, Withington, Manchester, United Kingdom. (1996-09)
      CD34+ cord blood cells were isolated with immunomagnetic beads and fractionated by fluorescence-activated cell sorting (FACS) into three subpopulations: CD34+38+DR+, CD34+38-DR+ and CD34+38-DR-, using antibodies specific for these cell surface markers. Cells from each of the three subsets were plated as single cells in serum-free medium supplemented with a combination of growth factor and individual cells were monitored for proliferation and the capacity to form colony-forming cells. Single cells from the CD34+38+DR+ subset showed the lowest expansion capacity, generating up to 1.1 x 10(6) cells at five weeks, while individual cells from both the CD34+38-DR+ and CD34+38-DR- subsets could be expanded up to 1.8 x 10(6) and 9.2 x 10(6) cells, respectively, over a period of six weeks. The different subpopulations also generated colony-forming cells which gave rise to erythroid, myeloid and erythroid/myeloid colonies. CD34+38-DR+ cells generated large numbers of colonies within two weeks in liquid culture, but this rapidly declined. Generation of lineage-committed colony-forming cells was better sustained in the CD34+38-DR- population and continued for up to six weeks in culture. Overall, the generation of colony-forming cells declined with time in culture, although the cell numbers continued to expand. However, when the same populations were plated on irradiated bone marrow stroma, both the CD34+38-DR+ and the CD34+38-DR- cells were capable of producing granulocytemacrophage colony-forming cells (GM-CFCs) for 10 to 12 weeks. As hemopoiesis was sustained for almost three months, it appears that these populations were significantly enriched in long-term culture-initiating cells (LTC-ICs). Although both populations generated GM-CFCs, the CD34+38-DR- cells sustained production of higher numbers of colony-forming cells than the CD34+38-DR+ population. These results demonstrate that cells from cord blood can be efficiently monitored at the single-cell level for proliferation, expansion and colony-forming capacity. Furthermore, at least two populations of LTC-ICs can be distinguished in cord blood CD34+38- cells by the differential expression of the HLA-DR antigen.
    • External quality assurance of circulating tumor cell enumeration using the CellSearch(®) system: a feasibility study.

      Kraan, J; Sleijfer, S; Strijbos, M; Ignatiadis, M; Peeters, D; Pierga, J; Farace, F; Riethdorf, S; Fehm, T; Zorzino, L; et al. (2011-03)
      Circulating tumor cells (CTCs) are cells that have detached from solid tumors and entered the blood. CTCs can be detected, among others, by semi-automated immunomagnetic enrichment and image cytometry using CellSearch® (Veridex, Raritan, NJ). We studied the feasibility of external quality assurance (EQA) of the entire CellSearch procedure from blood draw to interpretation of results in multiple laboratories.
    • External validation of survival of lung cancer patients due to setup uncertainties towards the heart

      Brink, C.; Bernchou, U.; Bertelsen, A.; Hansen, O.; Schytte, T.; Holloway, L.; Van Herk, Marcel; Johnson-Hart, Corinne; Price, Gareth J; Aznar, Marianne Camille; et al. (2020)
      Purpose or Objective The impact on survival due to heart toxicity for lung cancer patients treated with radiotherapy (RT) has been difficult to quantify, due to correlations between dose to heart and lung with tumor burden. A possible measure that indicates enhanced dose to heart but does not correlate with the other risk factors is the deviation between actual and planned daily distance between isocenter and heart. The average of this daily distance (DeltaD), which can be obtained from CBCT scans, will for positive values indicate larger separation of target and heart and thus reduced heart dose during delivery. DeltaD has previously been reported to impact survival [Johnson-Hart et al IJROBP 2018]. The aim of the current study is to undertake an external validation of this finding in another institution Material and Methods All patients treated in the validation department from April 2010 to end 2015 with daily CBCT, planned dose of 60-66 Gy in 2 Gy fractions and for which a heart delineation was available were collected for analysis. Standard clinical IGRT procedure utilized two registrations for each CBCT 1) tumor region (mask) and 2) overall anatomy (clipbox). The first was used for patient positioning while the latter was used for overall validation of the anatomy. The latter is representative of the heart position, while the first represents the tumor position hence the treatment isocenter. The difference between the registrations can thus be used to estimate the daily shift of the heart relative to the isocenter. Based on clinical data (performance status, GTV volume, Age, T and N stage, histology, tumor stage and dose) a base Cox model predicting survival was created using Lasso for parameter selection. Impact of continuous variable DeltaD was tested by adding DeltaD to the base Cox model. All analyses were performed in R v. 3.6.1. Results 300 patients were included. DeltaD did not correlate statistically significant with any of the clinical factors. Range of DeltaD was -0.6 cm to 0.7 cm. Figure 1 shows Kaplan-Meier plots of patients with positive versus negative DeltaD. It is seen that a split of the curves occurs around 16 months after the start of RT. The base Cox model included performance status, ln(GTV volume) and Age. DeltaD was included in the base model using time dependent Cox regression for which DeltaD effectively was zero until 16 months after RT. This resulted in a statistically significant regression constant of DeltaD of - 1.63 per cm (HR=0.195) with p-value of 0.017 Conclusion As in the original study this validation study demonstrates significant impact on survival due to estimated setup errors in the direction of the heart, even with daily online IGRT corrections. This indicates that dose to heart impacts survival for these patients. However, in contrast to the original study the effect of DeltaD only starts 16 months after RT. This difference in the two studies might reflect differences in treatment or patient cohorts at the two centers, and calls for additional external validation.
    • Extracellular matrix degradation and remodeling in development and disease.

      Lu, Pengfei; Takai, K; Weaver, V; Werb, Z; Breakthrough Breast Cancer Research Unit, Paterson Institute for Cancer Research and Wellcome Trust Centre for Cell Matrix Research, University of Manchester, United Kingdom. (2011-12)
      The extracellular matrix (ECM) serves diverse functions and is a major component of the cellular microenvironment. The ECM is a highly dynamic structure, constantly undergoing a remodeling process where ECM components are deposited, degraded, or otherwise modified. ECM dynamics are indispensible during restructuring of tissue architecture. ECM remodeling is an important mechanism whereby cell differentiation can be regulated, including processes such as the establishment and maintenance of stem cell niches, branching morphogenesis, angiogenesis, bone remodeling, and wound repair. In contrast, abnormal ECM dynamics lead to deregulated cell proliferation and invasion, failure of cell death, and loss of cell differentiation, resulting in congenital defects and pathological processes including tissue fibrosis and cancer. Understanding the mechanisms of ECM remodeling and its regulation, therefore, is essential for developing new therapeutic interventions for diseases and novel strategies for tissue engineering and regenerative medicine.
    • The extracellular matrix of human amniotic epithelium: ultrastructure, composition and deposition.

      Aplin, J; Campbell, S; Allen, Terence D; University of Manchester, St Mary's Hospital, Manchester M13 0JH, UK. (1985-11)
      Ultrastructural comparisons have been made between human amnion extracellular matrix in tissue and cell culture. Immunochemical analysis of matrix deposited by monolayers of cultured amnion epithelial cells has also been undertaken. The basal cell surfaces are highly invaginated with an associated basal lamina that is more electron dense at the distal tips of basal cell processes where hemidesmosomes are frequent. Immediately below the lamina densa is a zone rich in collagen bundles. In the underlying stroma two types of fibril predominate, one striated of 50 nm diameter and one of 18 nm diameter. The observations suggest that at gestational term the epithelial cells are still active in the production of matrix. Secretion appears to occur into invaginations in the basal cell surface where a loosely organized mixture of stromal-type and basal laminal-type aggregates is formed. In culture on plastic, cells also deposit a mixture of basal laminal (type IV collagen + laminin) and stromal (collagens type I + III) components as well as fibronectin. However, segregation into a true basal lamina with underlying stroma does not occur in vitro, suggesting the need for an organized subcellular template to complete matrix morphogenesis. The in vitro and in vivo evidence suggest that the epithelium contributes to the subjacent dense collagenous zone as well as to the basal lamina.
    • The extracellular matrix: a dynamic niche in cancer progression.

      Lu, Pengfei; Weaver, V; Werb, Z; Breakthrough Breast Cancer Research Unit, University of Manchester, Manchester M20 4BX, England, UK. (2012-02-20)
      The local microenvironment, or niche, of a cancer cell plays important roles in cancer development. A major component of the niche is the extracellular matrix (ECM), a complex network of macromolecules with distinctive physical, biochemical, and biomechanical properties. Although tightly controlled during embryonic development and organ homeostasis, the ECM is commonly deregulated and becomes disorganized in diseases such as cancer. Abnormal ECM affects cancer progression by directly promoting cellular transformation and metastasis. Importantly, however, ECM anomalies also deregulate behavior of stromal cells, facilitate tumor-associated angiogenesis and inflammation, and thus lead to generation of a tumorigenic microenvironment. Understanding how ECM composition and topography are maintained and how their deregulation influences cancer progression may help develop new therapeutic interventions by targeting the tumor niche.
    • Extrapolation from in vitro tests to human risk: experience with sodium fluoride clastogenicity.

      Scott, David; Roberts, Stephen A (1987-09)
      Genotoxic effects observed in vitro, only at high doses or high levels of cytotoxicity, will be false positives if such conditions are not achieved or cannot be tolerated in vivo. However, for such effects to be disregarded there must be a threshold dose or level of cytotoxicity below which genotoxicity is absent. Sodium fluoride (NaF) has previously been shown to be clastogenic in vitro in Syrian hamster cells and human fibroblasts. We have extended these studies in human fibroblasts and included a positive control (mitomycin C, MMC) which is clastogenic in vivo and carcinogenic, and a chemically related control (NaCl). Cytotoxicity was measured as mitotic inhibition and cell death (loss of clonogenicity). The results are used to illustrate the problems associated with quantitative extrapolation from in vitro tests to human risk, as follows. (1) There appears to be a threshold response (clastogenicity vs. dose) with NaF at around 10 micrograms/ml (48 h exposure) but a more definitive conclusion must await elucidation of the mechanisms of clastogenicity. (2) NaCl is weakly clastogenic at 1000 times the threshold dose for NaF. The mechanisms are unlikely to be similar. (3) No clastogenicity was detected with NaF below about 30% mitotic inhibition but the relationship between clastogenicity and mitotic inhibition was similar for NaF and MMC. (4) There was no obvious threshold in the relationship between clastogenicity and cell killing with NaF. MMC was less clastogenic than NaF at equitotoxic doses. Observations 3 and 4 preclude the possibility of regarding the clastogenicity of NaF as a false positive by virtue of associated cytotoxicity.
    • Extraskeletal myxoid chondrosarcoma with neuroendocrine differentiation: a pathologic, cytogenetic, and molecular study of a case with a novel translocation t(9;17)(q22;q11.2).

      Harris, Martin; Coyne, John; Tariq, M; Eyden, Brian P; Atkinson, M; Freemont, Anthony J; Varley, Jennifer; Attwooll, C; Telford, Nicholas; Department of Histopathology, Christie Hospital, Manchester, UK. (2000-07)
      A case of extraskeletal myxoid chondrosarcoma (EMC) in which there was histochemical, immunohistochemical, and ultrastructural evidence of neuroendocrine differentiation is reported. Genetic investigations showed the recently described novel translocation t(9;17)(q22;q11.2) and associated fusion of the CHN and RBP56 genes, contrasting with the translocation t(9;22)(q22;q12) and EWS/CHN gene fusion found in the majority of EMCs.
    • Extravascular natural cytotoxicity in man: Anti-K562 activity of lymph-node and tumour-infiltrating lymphocytes.

      Moore, Michael; Vose, Brent M; The Paterson Laboratories, Christie Hospital and Holt Radium Institute, Withington, Manchester M20 9BX (1981-03-15)
      A major distinction is reported between the cytolytic activity of peripheral blood lymphocytes (PBL) and that of lymph-node and tumour-infiltrating lymphocytes (TIL) against targets of the NK-sensitive K562 cell line in short-term 51Cr release assays. In the PBL of normal donors and lung cancer patients in whom disease was advanced anti-K562 reactivity, though variable, was consistently detectable and this activity could be augmented to a similar extent in patients and controls by treatment of effectors with exogenous interferon (IF). By contrast anti-K562 activity in lymph-node cells (LNC) and TIL was virtually absent and significant levels could not be induced by exposure to IF. This activity was not attributable to coexistent suppressor cells for NK function since admixture of LNC and TIL with normal PBL failed to modulate K562 killing by the latter. The results imply that K562-reactive NK cells and their precursors may frequently be present at sub-threshold levels in the lymph nodes of tumour-bearing patients and a similar explanation could account for the inactivity of TIL. However, in the latter situation, actively-induced NK dysfunction in situ incapable of regeneration by IF, and attributable to as yet undefined tumour-associated factors, may also be involved.
    • Extreme incidence of skin cancer in kidney and liver transplant recipients living with high sun exposure

      Plasmeijer, EI; Jiyad, Z; Way, M; Marquart, L; Miura, K; Campbell, S; Isbel, N; Fawcett, J; Ferguson, LE; Davis, M; et al. (2019)
    • Extreme sensitivity of some intestinal crypt cells to X and gamma irradiation.

      Potten, Christopher S; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester (1977-10-06)
    • Ezh2 is essential for the generation of functional yolk sac derived erythro-myeloid progenitors

      Neo, Wen H; Meng, Y.; Rodriguez-Meira, A.; Fadlullah, Muhammad Z H; Booth, C. A. G.; Azzoni, E.; Thongjuea, S.; de Bruijn, M.; Jacobsen, S. E. W.; Mead, A. J.; et al. (2021)
    • FACT: An Open-Label Randomized Phase III Study of Fulvestrant and Anastrozole in Combination Compared With Anastrozole Alone as First-Line Therapy for Patients With Receptor-Positive Postmenopausal Breast Cancer.

      Bergh, Jonas; Jönsson, P; Lidbrink, E; Trudeau, M; Eiermann, W; Brattström, D; Lindemann, J; Wiklund, F; Henriksson, R; Manchester University and Christie Hospital, Manchester, United Kingdom. (2012-03-12)
      PURPOSE To compare the effect of therapy with anastrozole versus a combination of fulvestrant and anastrozole in women in first relapse of endocrine-responsive breast cancer. PATIENTS AND METHODS Postmenopausal women, or premenopausal women receiving a gonadotropin-releasing hormone agonist, with estrogen receptor- and/or progesterone receptor-positive disease at first relapse after primary treatment of localized disease were open-label randomly assigned to a fulvestrant loading dose (LD) regimen followed by monthly injection plus 1 mg of anastrozole daily or to 1 mg of anastrozole daily alone. The primary end point was time to progression (TTP). RESULTS: 63 patients (24.6%) versus 35 patients (13.8%) in the standard arm (P = .0023). Death owing to AEs was reported in 11 (4.3%) and five patients (2.0%) in the experimental versus standard arm, respectively. CONCLUSION Fulvestrant (250 mg + LD regimen) in combination with anastrozole offered no clinical efficacy advantage over anastrozole monotherapy in this population of individuals with a relatively high proportion of previous adjuvant antiestrogen exposure.
    • Factors affecting the growth of Chinese hamster cells in HAT selection media.

      Fox, Margaret; Boyle, John M (1976-06)
      Factors affecting the efficiency of selection of "revertants" of salvage pathway mutants in media containing amethopterin have been examined. Our V79 Chinese hamster cell line was found to require a significantly higher level of thymidine for optimal growth in such media than has been reported for other cell lines. Hypoxanthine (but not glycine) was also required for reversal of amethopterin toxicity, but levels did not differ significantly from those reported elsewhere. Growth in HAT was also dependent on plating density and serum batch. Our modification (VHAT) was compared with published HAT recipies in back selection reconstruction experiments. A sharp fall in EOR (efficiency of recovery) of wild type cells from mixtures with mutants at plating densities greater than 3500 cells/cm2 (10(5) cells/6 cm dish) was observed for VHAT. EOR with other HAT recipes was lower still, and was affected also by the particular mutant used in the mixture. EMS induced "revertants" were isolated from three 8AZr mutants by plating in VHAT. All revertants were however amethopterin resistant, they were also 8AZ resistant and the mobility of residual HGPRT (as measured by polyacrylamide gel electrophoresis) was similar to that of their 8AZr parents i.e. dissimilar from that in wild type. The modal chromosome number of V79 wild type cells was 21. No significant deviation from this mode was detected in any of the mutant lines examined. The data indicate that the recovery of colonies in HAT from 8AZr mutants does not necessarily indicate that a back mutation in the structural gene for HGPRT has occurred. Thus, the frequency of HAT+ colonies cannot be taken as a direct indication of reversion frequencies.
    • Factors affecting the growth of Chinese hamster cells in HAT selection media.

      Fox, Margaret; Boyle, John M; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester (1976-06)
      Factors affecting the efficiency of selection of "revertants" of salvage pathway mutants in media containing amethopterin have been examined. Our V79 Chinese hamster cell line was found to require a significantly higher level of thymidine for optimal growth in such media than has been reported for other cell lines. Hypoxanthine (but not glycine) was also required for reversal of amethopterin toxicity, but levels did not differ significantly from those reported elsewhere. Growth in HAT was also dependent on plating density and serum batch. Our modification (VHAT) was compared with published HAT recipies in back selection reconstruction experiments. A sharp fall in EOR (efficiency of recovery) of wild type cells from mixtures with mutants at plating densities greater than 3500 cells/cm2 (10(5) cells/6 cm dish) was observed for VHAT. EOR with other HAT recipes was lower still, and was affected also by the particular mutant used in the mixture. EMS induced "revertants" were isolated from three 8AZr mutants by plating in VHAT. All revertants were however amethopterin resistant, they were also 8AZ resistant and the mobility of residual HGPRT (as measured by polyacrylamide gel electrophoresis) was similar to that of their 8AZr parents i.e. dissimilar from that in wild type. The modal chromosome number of V79 wild type cells was 21. No significant deviation from this mode was detected in any of the mutant lines examined. The data indicate that the recovery of colonies in HAT from 8AZr mutants does not necessarily indicate that a back mutation in the structural gene for HGPRT has occurred. Thus, the frequency of HAT+ colonies cannot be taken as a direct indication of reversion frequencies.
    • Factors affecting the quantitation of dose-response curves for mutation induction in V79 Chinese hamster cells after exposure to chemical and physical mutagens.

      Fox, Margaret; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester (1975-09)
      Using four common mutagens, ethyl methanesulphonate (EMS), methyl methanesulphonate (mms), uv, and X-irradiation, the relationship between dose of mutagen, cellular lethality and frequency of 8-azaguanine resistant colonies in V79 Chinese hamster cells has been examined. Several factors affecting the recovery of mutants including inter and intra-clone metabolic co-operation have been quantitated and their influence on survival response curves examined. Induced mutant frequencies were assayed by two methods in situ, and after replating. After exposure to X-rays, MMS and UV a significantly higher frequency of mutants was observed in replated experiments as compared with the in situ situation, at all survival levels assayed. With EMS, an increment on replating was observed only at high survival levels. The replating data suggest that two types of azgr colonies are produced, i.e. those which contain only azgr cells and those which, due to damage segregation, contain a mixture of azgr and azg8 cells. These mixed colonies appear to be lost by metabolic co-operation when mutation frequencies are assayed in silu. The proportion of mixed to homogeneous colonies differs with different mutagens. Taking into account such factors, EMS and UV irradiation were similarly mutagenic at a given survival level, but at equitoxic doses, fewer mutants were recovered after exposure of V79 cells to MMS and X-rays.
    • Factors influencing the yield of free radicals in irradiated chicken bones

      Dodd, Nicholas J F; Haishun, J; Lea, J S; Swallow, A John; Paterson Institute for Cancer Research, Christie Hospital and Holt Radium Institute, Manchester, UK (1992)