• Prostate cancer evolution from multilineage primary to single lineage metastases with implications for liquid biopsy

      Woodcock, D. J.; Riabchenko, E.; Taavitsainen, S.; Kankainen, M.; Gundem, G.; Brewer, D. S.; Ellonen, P.; Lepistö, M.; Golubeva, Y. A.; Warner, A. C.; et al. (2020)
      The evolutionary progression from primary to metastatic prostate cancer is largely uncharted, and the implications for liquid biopsy are unexplored. We infer detailed reconstructions of tumor phylogenies in ten prostate cancer patients with fatal disease, and investigate them in conjunction with histopathology and tumor DNA extracted from blood and cerebrospinal fluid. Substantial evolution occurs within the prostate, resulting in branching into multiple spatially intermixed lineages. One dominant lineage emerges that initiates and drives systemic metastasis, where polyclonal seeding between sites is common. Routes to metastasis differ between patients, and likely genetic drivers of metastasis distinguish the metastatic lineage from the lineage that remains confined to the prostate within each patient. Body fluids capture features of the dominant lineage, and subclonal expansions that occur in the metastatic phase are non-uniformly represented. Cerebrospinal fluid analysis reveals lineages not detected in blood-borne DNA, suggesting possible clinical utility.
    • Prostate cancer heterogeneity assessment with multi-regional sampling and alignment-free methods

      Murphy, R. G.; Roddy, A. C.; Srivastava, S.; Baena, Esther; Waugh, D. J.; J, M. O. S.; McArt, D. G.; Jain, S.; LaBonte, M. J.; Movember FASTMAN Centre of Excellence, Patrick G Johnston Centre for Cancer Research, School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast BT9 7AE, UK. (2020)
      Combining alignment-free methods for phylogenetic analysis with multi-regional sampling using next-generation sequencing can provide an assessment of intra-patient tumour heterogeneity. From multi-regional sampling divergent branching, we validated two different lesions within a patient's prostate. Where multi-regional sampling has not been used, a single sample from one of these areas could misguide as to which drugs or therapies would best benefit this patient, due to the fact these tumours appear to be genetically different. This application has the power to render, in a fraction of the time used by other approaches, intra-patient heterogeneity and decipher aberrant biomarkers. Another alignment-free method for calling single-nucleotide variants from raw next-generation sequencing samples has determined possible variants and genomic locations that may be able to characterize the differences between the two main branching patterns. Alignment-free approaches have been applied to relevant clinical multi-regional samples and may be considered as a valuable option for comparing and determining heterogeneity to help deliver personalized medicine through more robust efforts in identifying targetable pathways and therapeutic strategies. Our study highlights the application these tools could have on patient-aligned treatment indications.
    • A prostate cancer proteomics database for SWATH-MS based protein quantification

      Muazzam, Ammara; Chiasserini, D.; Kelsall, J.; Geifman, N.; Whetton, Anthony D; Townsend, Paul A; Manchester Cancer Research Centre, Division of Cancer Sciences, Faculty of Biology, School of Medical Sciences, Medicine and Health, University of Manchester, Wilmslow Road, Manchester M20 4GJ, UK (2021)
      Prostate cancer is the most frequent form of cancer in men, accounting for more than one-third of all cases. Current screening techniques, such as PSA testing used in conjunction with routine procedures, lead to unnecessary biopsies and the discovery of low-risk tumours, resulting in overdiagnosis. SWATH-MS is a well-established data-independent (DI) method requiring prior knowledge of targeted peptides to obtain valuable information from SWATH maps. In response to the growing need to identify and characterise protein biomarkers for prostate cancer, this study explored a spectrum source for targeted proteome analysis of blood samples. We created a comprehensive prostate cancer serum spectral library by combining data-dependent acquisition (DDA) MS raw files from 504 patients with low, intermediate, or high-grade prostate cancer and healthy controls, as well as 304 prostate cancer-related protein in silico assays. The spectral library contains 114,684 transitions, which equates to 18,479 peptides translated into 1227 proteins. The robustness and accuracy of the spectral library were assessed to boost confidence in the identification and quantification of prostate cancer-related proteins across an independent cohort, resulting in the identification of 404 proteins. This unique database can facilitate researchers to investigate prostate cancer protein biomarkers in blood samples. In the real-world use of the spectrum library for biomarker detection, using a signature of 17 proteins, a clear distinction between the validation cohort's pre- and post-treatment groups was observed. Data are available via ProteomeXchange with identifier PXD028651.
    • Prostate cancer radiation therapy recommendations in response to COVID-19

      Zaorsky, N. G.; Yu, J. B.; McBride, S. M.; Dess, R. T.; Jackson, W. C.; Mahal, B. A.; Chen, R.; Choudhury, Ananya; Henry, A.; Syndikus, I.; et al. (2020)
      Purpose: During a global pandemic, the benefit of routine visits and treatment of patients with cancer must be weighed against the risks to patients, staff, and society. Prostate cancer is one of the most common cancers radiation oncology departments treat, and efficient resource utilization is essential in the setting of a pandemic. Herein, we aim to establish recommendations and a framework by which to evaluate prostate radiation therapy management decisions. Methods and materials: Radiation oncologists from the United States and the United Kingdom rapidly conducted a systematic review and agreed upon recommendations to safely manage patients with prostate cancer during the COVID-19 pandemic. A RADS framework was created: remote visits, and avoidance, deferment, and shortening of radiation therapy was applied to determine appropriate approaches. Results: Recommendations were provided by the National Comprehensive Cancer Network risk group regarding clinical node-positive, postprostatectomy, oligometastatic, and low-volume M1 disease. Across all prostate cancer stages, telemedicine consultations and return visits were recommended when resources/staff available. Delays in consultations and return visits of between 1 and 6 months were deemed safe based on stage of disease. Treatment can be avoided or delayed until safe for very low, low, and favorable intermediate-risk disease. Unfavorable intermediate-risk, high-risk, clinical node-positive, recurrence postsurgery, oligometastatic, and low-volume M1 disease can receive neoadjuvant hormone therapy for 4 to 6 months as necessary. Ultrahypofractionation is preferred for localized, oligometastatic, and low-volume M1, and moderate hypofractionation is preferred for postprostatectomy and clinical node positive disease. Salvage is preferred to adjuvant radiation. Conclusions: Resources can be reduced for all identified stages of prostate cancer. The RADS (remote visits, and avoidance, deferment, and shortening of radiation therapy) framework can be applied to other disease sites to help with decision making in a global pandemic.
    • Prostate radiotherapy with carbogen and nicotinamide. Final results of the phase 1b/II PROCON trial

      Yip, K.; Hoskin, Peter J; Thiruthaneeswaran, Niluja; Alonzi, R.; Marie Curie Research Wing, Mount Vernon Cancer Centre, Northwood Middlesex, (2020)
      Purpose or Objective Prostate tumour hypoxia is associated with resistance to radiotherapy (RT) and increased likelihood of relapse post treatment. The concurrent administration of carbogen and nicotinamide (CON) with RT improves overall survival in patients with bladder cancer and regional control rates following laryngeal RT. We evaluated the safety, toxicity of this approach for patients with high risk prostate cancer. Material and Methods 50 patients (with 1 of: T3, Gleason ≥8, PSA ≥20) were recruited into the single arm phase 1b/II PROCON trial between Dec 2011 and Sept 2013. They breathed carbogen via a tight-fitting face mask during RT and took 60mg/kg of nicotinamide daily prior to the delivery of RT (74Gy/37# to the prostate and 60Gy/37# to the pelvic nodes). Twenty men also underwent functional MRI imaging; correction of tumour hypoxia was evaluated by comparing tumour R2* values before and after carbogen administration. Patients were assessed during treatment and 2, 4 and 12 weeks after they had completed RT. The primary endpoints were prevalence of grade 3 (G3) or worse lower gastrointestinal (GI) or genitourinary (GU) toxicities during this period. Secondary end points include PSA progression free survival (PFS) and overall survival (OS) rates at five years as calculated by the Kaplan Meier method. Results The median duration of follow up was 60 months. All patients completed the prescribed 37 fraction schedule of RT over 50 to 56 days (median duration: 52 days). 56% patients experienced grade 1 nausea (CTCAE v 4.0), and 52% patients received anti-emetic treatment. 22% patients had a nicotinamide dose adjustment, 20% patients had a break from nicotinamide and 6% patients discontinued nicotinamide. 96% patients received carbogen with every RT fraction. One patient had an interruption of treatment and received carbogen for 33 of 37 fractions. One patient discontinued carbogen treatment after 7 fractions. No one developed G3 or worse lower GI or GU toxicities; prevalence of G1/G2 toxicities are shown in table 1. The 5 year OS and PSA-PFS rates were 92% and 87% respectively. Six patients experienced biochemical progression. Five patients have died but None had biochemical relapse prior to their death. The functional MRI analysis showed a mean decrease of 5.8% in tumour R2* after the application of carbogen, measured in 72 pairs of pre and post carbogen measurements, indicating that the carbogen exposure resulted in an immediate reduction in tumour hypoxia. Conclusion The prevalence of acute lower GI and GU toxicities among our cohort, together with their 5 year PSA PFS and OS rates, are comparable, or superior to, those reported for patients with high risk prostate cancer receiving RT in other contemporary trials. The concurrent administration of CON with RT is safe and effective. It should be further explored in a phase III setting.
    • Protection against mucosal injury by growth factors and cytokines.

      Booth, Dawn; Potten, Christopher S; Cancer Research Campaign Epithelial Biology Group, Paterson Institute for Cancer Research, Christie Hospital, Manchester, UK. (2001)
      This article provides an overview of published studies in which growth factors and cytokines were used to modify the sensitivity of intestinal stem cells to a dose of radiation. In these experiments, growth factors were used to manipulate the sensitivity of stem cells in the gastrointestinal tract to reduce the severity of gastrointestinal mucositis in cancer therapy patients. Transforming growth factor beta3, interleukin 11, and keratinocyte growth factor were used. All three agents, given according to appropriate protocols, can result in a threefold to fourfold increase in the number of intestinal stem cells that survive a dose of radiation therapy. This result was assessed by using the crypt microcolony assay of stem cell functional capacity. The changes in stem cell survival that were observed resulted in increased animal survival. This increased survival was taken as a surrogate for improvement in patient well-being. The severity of diarrhea, a marker of functional impairment, was concomitantly reduced.
    • Protection and selection for gene therapy in the hematopoietic system.

      Milsom, Michael D; Fairbairn, Leslie J; Cancer Research UK Gene Therapy Group, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Wilmslow Road, Manchester M20 4BX, UK. (2004-02)
      Hematopoietic stem cell gene therapy is potentially curative for a number of inherited and acquired disorders. However, poor gene transfer and expression in repopulating hematopoietic stem cells attenuate this potential. Here we review potential means of conferring a selective advantage to hematopoietic stem cells and their progeny, and discuss the issues that surround the use of selective advantages in vivo.
    • Protection by ultraviolet A and B sunscreens against in situ dipyrimidine photolesions in human epidermis is comparable to protection against sunburn.

      Young, Antony R; Sheehan, John M; Chadwick, Caroline A; Potten, Christopher S; Department of Environmental Dermatology, St. John's Institute of Dermatology, King's College London, St. Thomas's Hospital, London, UK. antony.r.young@kcl.ac.uk (2000-07)
      Sunscreens prevent sunburn and may also prevent skin cancer by protecting from ultraviolet-induced DNA damage. We assessed the ability of two sunscreens, with different spectral profiles, to inhibit DNA photodamage in human epidermis in situ. One formulation contained the established ultraviolet B filter octyl methoxycinnamate, whereas the other contained terephthalylidene dicamphor sulfonic acid, a new ultraviolet A filter. Both formulations had sun protection factors of 4 when assessed with solar simulating radiation in volunteers of skin type I/II. We tested the hypothesis that sun protection factors would indicate the level of protection against DNA photodamage. Thus, we exposed sunscreen-treated sites to four times the minimal erythema dose of solar simulating radiation, whereas vehicle and control sites were exposed to one minimal erythema dose. We used monoclonal antibodies against thymine dimers and 6-4 photoproducts and image analysis to quantify DNA damage in skin sections. A dose of four times the minimal erythema dose, with either sunscreen, resulted in comparable levels of thymine dimers and 6-4 photoproducts to one minimal erythema dose +/- vehicle, providing evidence that the DNA protection factor is comparable to the sun protection factor. The lack of difference between the sunscreens indicates similar action spectra for erythema and DNA photodamage and that erythema is a clinical surrogate for DNA photodamage that may lead to skin cancer.
    • Protection of Chinese hamster cells against the cytotoxic and mutagenic effects of alkylating agents by transfection of the Escherichia coli alkyltransferase gene and a truncated derivative.

      Fox, Margaret; Brennand, J; Margison, Geoffrey P; Department of Biochemical Genetics, Paterson Institute for Cancer Research, Withington, Manchester, UK. (1987-11)
      The cytotoxic and mutagenic effects of various monofunctional and bifunctional alkylating agents have been assessed in V79 Chinese hamster cells that express either the entire O6-alkylguanine (O6AG) and alkylphosphotriester alkyltransferase (ATase) gene (clone 8 cells) or a truncated form that codes only for O6AG ATase activity (clone SB cells). Protection ratios, as determined by D37 values, were greater for clone 8 cells than for SB cells. Significant protection against the mutagenic effects of N-methyl-N-nitrosourea and ethylmethanesulphonate at the hypoxanthine phosphoribosyltransferase (HPRT) locus was observed in clone 8 and SB cells. Streptozotocin and the haloethyl nitrosoureas, chlorozotocin and bis-chloroethylnitrosourea were less efficient in inducing HPRT-deficient mutants and a smaller degree of protection was afforded by the transfected genes. This is possibly due to the propensity of these compounds to induce multi-locus deletions. Southern analysis of DNA from clone 8 and SB cells indicated the presence of multiple copies of the plasmid integrated into clone 8 cells but few copies in clone SB cells. The copy number did not change but ATase levels fell when cells were grown in the absence of G418.
    • Protection of committed murine haemopoietic progenitors against BCNU toxicity does not predict protection of primitive, multipotent spleen colony-forming cells - implications for chemoprotective gene therapy.

      Chinnasamy, Nachimuthu; Rafferty, Joseph A; Lashford, Linda S; Chinnasamy, Dhanalakshmi; Margison, Geoffrey P; Thatcher, Nick; Dexter, T Michael; Fairbairn, Leslie J; CRC Sections of Genome Damage and Repair, Paterson Institute for Cancer Research, Manchester, UK. (1999-11)
      The effect of expression of an O6-benzylguanine (O6-beG)-resistant mutant (hATPA/GA) of human O6-alkylguanine-DNA alkyltransferase (ATase) on the in vivo toxicity and clastogenicity of the anti-tumour agent N,N'-bis(2-chloroethyl)-N-nitrosourea (BCNU) to murine bone marrow has been investigated. When compared with control animals, the bipotent granulocyte-macrophage colony-forming (GM-CFC) progenitor population of the hATPA/GA transduced mice were somewhat more resistant to BCNU (1.4-fold, P = 0.047) and this effect was more significant in the presence of the ATase inactivator O6-beG (3. 5-fold, P = 0.001). The polychromatic erythrocytes were also significantly protected against BCNU-induced clastogenicity both in the presence (P < 0.001) and absence of O6-beG (P < 0.05). The primitive, multipotent spleen colony-forming cells (CFU-S) in these animals also showed moderate (1.6-fold, P = 0.034) protection in the absence of O6-beG but in the presence of the inactivator they remained as sensitive to BCNU toxicity as those in the control animals (P = 0.133). This result contrasts with previous findings demonstrating significant hATPA/GA-mediated, O6-beG-resistant protection against the toxicity and clastogenicity of a number of O6-alkylating agents, including temozolomide, fotemustine and chlorozotocin. The possibility that our strategy for protective gene therapy may be highly agent and cell-type specific is unexpected and has possible implications for clinical trials of this approach using BCNU or related agents.
    • Protection of mammalian cells against chloroethylating agent toxicity by an O6-benzylguanine-resistant mutant of human O6-alkylguanine-DNA alkyltransferase.

      Hickson, Ian; Fairbairn, Leslie J; Chinnasamy, Nachimuthu; Dexter, T Michael; Margison, Geoffrey P; Rafferty, Joseph A; CRC Department of Experimental Haematology, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, UK. (1996-10)
      Low levels of expression in haemopoietic cells of the DNA repair protein O6-alkylguanine-DNA alkyltransferase (A Tase), is associated with the dose-limiting sensitivity of these cells to the chemotherapeutic chloroethylating and related methylating agents. Thus, the use of agents which deplete ATase such as O6-benzylguanine (O6-beG), as a tumour sensitisation strategy is likely further to potentiate collateral toxicity in bone marrow. In order to address this problem, we have engineered two mutants of human ATase (hAT) for resistance to O6-beG and characterised the in vitro properties of the proteins. In one mutant protein (hATPA), the proline at position 140 was changed to an alanine, whilst in the other (hATPA/GA) an additional mutation (glycine 156 to alanine) was also introduced. The I50 values for O6-beG of hAT, hATPA and hATPA/GA are 0.16, 2.5 and > 500 microM respectively, indicating that hATPA is resistant and hATPA/GA effectively refractory to O6-beG inactivation. Both mutant proteins retain comparable methyl transfer kinetics to those of nonmutant hAT and although they are thermally less stable in vitro than the wild-type protein, both can be substantially stabilised by DNA. Expression of either hAT or hATPA/GA following gene transfer into RJKO cells, raised the D37 value for mitozolomide from 0.35 microgram/ml for control cells to 10 micrograms/ml in the absence of O6-beG. However, whilst hAT-mediated protection was ablated by 20 microM O6-beG, the hATPA/GA protein provided protection against mitozolomide under the same conditions. Similar observations were made with chlorozotocin. The data suggest that transfer and expression of O6-beG resistant ATase in normal progenitor cells, should be a useful therapeutic strategy to protect the cells from the cytotoxic effects of the O6-alkylating agents even when used in combination with tumour sensitising agents such as O6-beG.
    • Protection of the small intestinal clonogenic stem cells from radiation-induced damage by pretreatment with interleukin 11 also increases murine survival time.

      Potten, Christopher S; CRC Department of Epithelial Biology, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, United Kingdom. (1996-07)
      The effect of administering recombinant human interleukin 11 in conjunction with cytotoxic insults to the gastrointestinal tract has been studied using the crypt microcolony assay for stem cell function and whole-animal survival time studies. The cytotoxic regimens include single doses of gamma rays; single doses of 5-fluorouracil (5-FU) and multiple doses of 5-FU spaced 6 h apart. Interleukin 11 (IL-11) (100 micrograms/kg) delivered over a period of time prior to cytotoxic exposure afforded protection to the clonogenic cells in the crypts as seen with the microcolony assay and prolonged the animal survival time following radiation exposure. Continuing this dose of IL-11 after cytotoxic exposure afforded little additional protection. Three doses of 5-FU 6 h apart generated crypt survival curves similar to those obtained after a single dose of gamma rays. IL-11 given prior to two doses of 5-FU effectively abolished the cytotoxic effect of the second dose of 5-FU; i.e., 2.5-3.0 times more crypts survived if IL-11 was administered when the higher 5-FU doses are considered. IL-11 given before a dose of 12 Gy of gamma rays prolonged the survival time of animals by three to four days. This confirms earlier studies demonstrating that protecting clonogenic cells in the crypt survival assay can result in beneficial effects on whole-animal survival times.
    • Protective immunization against Epstein-Barr virus-induced disease in cottontop tamarins using the virus envelope glycoprotein gp340 produced from a bovine papillomavirus expression vector.

      Finerty, S; Tarlton, J; Mackett, Mike; Conway, Margaret J; Arrand, John R; Watkins, P E; Morgan, A J; Department of Pathology, School of Medical Sciences, University of Bristol, U.K. (1992-02)
      Inoculation with Epstein-Barr virus (EBV) induces malignant lymphomas in the cottontop tamarin (Saguinus oedipus oedipus). This provides an experimental animal model for assessing the efficacy of candidate EBV vaccines which are intended to reduce the incidence of human tumours associated with EBV infection. Previous work has shown that experimental vaccines based on the major virus envelope glycoprotein gp340 prepared from the membranes of EBV-infected cells are effective in protecting cottontop tamarins against EBV-induced disease. However, not all purified gp340 preparations induce protective immunity against EBV lymphoma in the tamarin. In this work, cottontop tamarins were immunized with recombinant gp340, produced using a bovine papillomavirus (BPV) expression vector, and a threonyl muramyl dipeptide adjuvant formulation. Although the recombinant-derived gp340 lacked the membrane anchor sequence of authentic gp340 and was expressed in mouse cells, it was immunogenic and induced virus-neutralizing antibodies. Healthy vaccinated tamarins were protected against EBV-induced disease. The demonstration that a recombinant gp340 product is able to elicit protective immunity in the cottontop tamarin is a significant step in the development of an EBV vaccine because previously it had not been clear whether a recombinant product would have the exact tertiary structure, including the necessary carbohydrate components, to induce protective immunity. A recombinant gp340 vaccine offers various advantages over production of the authentic molecule by laborious biochemical separation, including lower cost and the absence of potentially oncogenic EBV DNA. Therefore, recombinant gp340 produced using the BPV expression vector is suitable for development as a candidate EBV vaccine for a human Phase I trial and beyond.
    • Protein kinase C activators can interact synergistically with granulocyte colony-stimulating factor or interleukin-6 to stimulate colony formation from enriched granulocyte-macrophage colony-forming cells.

      Heyworth, Clare M; Dexter, T Michael; Nicholls, Sian E; Whetton, Anthony D; Department of Experimental Haematology, Paterson Laboratories, Christie Hospital NHS Trust, Withington, Manchester, UK. (1993-02-15)
      The effects of direct activators of protein kinase C (PKC) (the phorbol ester tetradecanoyl phorbol myristic acid [TPA] or bryostatin) on the ability of a highly enriched population of granulocyte-macrophage colony-forming cells (GM-CFC) to proliferate and develop in soft agar was assessed. In the absence of colony stimulating factors, the PKC activators did not stimulate colony formation. However, in the presence of optimal concentrations of granulocyte colony-stimulating factor (G-CSF) or interleukin-6 (IL-6), TPA or bryostatin markedly elevated the number of colonies formed from the GM-CFC. In the absence of TPA, IL-6, and G-CSF, respectively, both stimulated the formation of about 3% of the colonies observed when IL-3 was present. When TPA plus G-CSF or IL-6 were added together, this figure increased to 48% and 54%, respectively. In both instances, the types of mature cells formed was altered from colonies of mature neutrophilic cells to a mixture consisting predominantly of macrophages with some neutrophils. Similar results were observed when bryostatin replaced TPA in these assays. When single cell colony-forming assays were performed, the same results were obtained. The presence of G-CSF, or IL-6, and the activator of PKC used (TPA or bryostatin) was required throughout the colony-forming assay for an optimal synergistic effect to be observed. These data indicate that agents that activate PKC can promote the proliferation and development of GM-CFC via a synergistic interaction with G-CSF or IL-6. Furthermore, there is an apparent role for PKC in development and possibly lineage commitment of GM-CFC.
    • Protein kinase C delta is phosphorylated on five novel Ser/Thr sites following inducible overexpression in human colorectal cancer cells

      Welman, Arkadiusz; Griffiths, John R; Whetton, Anthony D; Dive, Caroline; Cancer Research UK, Clinical and Experimental Pharmacology Group, Paterson Institute for Cancer Research, University of Manchester, Manchester M20 4BX, United Kingdom. awelman@picr.man.ac.uk (2007-12)
      Phosphorylation plays an important role in regulation of protein kinase C delta (PKCdelta). To date, three Ser/Thr residues (Thr 505, Ser 643, and Ser 662) and nine tyrosine residues (Tyr 52, Tyr 64, Tyr 155, Tyr 187, Tyr 311, Tyr 332, Tyr 512, Tyr 523, and Tyr 565) have been defined as regulatory phosphorylation sites for this protein (rat PKCdelta numbering). We combined doxycycline-regulated inducible gene expression technology with a hypothesis-driven mass spectrometry approach to study PKCdelta phosphorylation pattern in colorectal cancer cells. We report identification of five novel Ser/Thr phosphorylation sites: Thr 50, Thr 141, Ser 304, Thr 451, and Ser 506 (human PKCdelta numbering) following overexpression of PKCdelta in HCT116 human colon carcinoma cells grown in standard tissue culture conditions. Identification of potential novel phosphorylation sites will affect further functional studies of this protein, and may introduce additional complexity to PKCdelta signaling.
    • Protein kinase cδ deficiency causes mendelian systemic lupus erythematosus with B cell-defective apoptosis and hyperproliferation.

      Belot, A; Kasher, P; Trotter, Eleanor W; Foray, A-P; Debaud, A-L; Rice, G; Szynkiewicz, M; Zabot, M-T; Rouvet, I; Bhaskar, S; et al. (2013-08)
      Systemic lupus erythematosus (SLE) is a prototype autoimmune disease that is assumed to occur via a complex interplay of environmental and genetic factors. Rare causes of monogenic SLE have been described, providing unique insights into fundamental mechanisms of immune tolerance. The aim of this study was to identify the cause of an autosomal-recessive form of SLE.
    • Protein kinase signaling networks in cancer.

      Brognard, J; Hunter, Tony; Signalling Networks in Cancer Group, Cancer Research UK, Paterson Institute for Cancer Research, The University of Manchester, Manchester, UK. (2010-11-29)
      Protein kinases orchestrate the activation of signaling cascades in response to extracellular and intracellular stimuli to control cell growth, proliferation, and survival. The complexity of numerous intracellular signaling pathways is highlighted by the number of kinases encoded by the human genome (539) and the plethora of phosphorylation sites identified in phosphoproteomic studies. Perturbation of these signaling networks by mutations or abnormal protein expression underlies the cause of many diseases including cancer. Recent RNAi screens and cancer genomic sequencing studies have revealed that many more kinases than anticipated contribute to tumorigenesis and are potential targets for inhibitor drug development intervention. This review will highlight recent insights into known pathways essential for tumorigenesis and discuss exciting new pathways for therapeutic intervention.
    • Protein Z: a putative novel biomarker for early detection of ovarian cancer

      Russell, M; Walker, M; Williamson, A; Gentry-Maharaj, A; Ryan, A; Kalsi, J; Skates, S; D'Amato, A; Dive, Caroline; Pernemalm, M; et al. (2016-01-27)
      Ovarian cancer (OC) has the highest mortality of all gynaecological cancers. Early diagnosis offers an approach to achieving better outcomes. We conducted a blinded-evaluation of prospectively collected preclinical serum from participants in the multimodal group of the United Kingdom Collaborative Trial of Ovarian Cancer Screening. Using isobaric tags (iTRAQ) we identified 90 proteins differentially expressed between OC cases and controls. A second targeted mass spectrometry analysis of twenty of these candidates identified Protein Z as a potential early detection biomarker for OC. This was further validated by ELISA analysis in 482 serial serum samples, from 80 individuals, 49 OC cases and 31 controls, spanning up to 7 years prior to diagnosis. Protein Z was significantly down-regulated up to 2 years pre-diagnosis (p = 0.000000411) in 8 of 19 Type I patients whilst in 5 Type II individuals, it was significantly up-regulated up to 4 years before diagnosis (p=0.01). ROC curve analysis for CA-125 and CA-125 combined with Protein Z showed a statistically significant (p= 0.00033) increase in the AUC from 77% to 81% for Type I and a statistically significant (p= 0.00003) increase in the AUC from 76% to 82% for Type II. Protein Z is a novel independent early detection biomarker for Type I and Type II ovarian cancer; which can discriminate between both types. Protein Z also adds to CA-125 and potentially the Risk of Ovarian Cancer algorithm in the detection of both subtypes. This article is protected by copyright. All rights reserved. © 2014 Wiley Periodicals, Inc.
    • Proteoglycan-specific molecular switch for RPTPσ clustering and neuronal extension.

      Coles, Charlotte H; Shen, Yingjie; Tenney, Alan P; Siebold, Christian; Sutton, Geoffrey C; Lu, Weixian; Gallagher, John T; Jones, E Yvonne; Flanagan, John G; Aricescu, A Radu; et al. (2011-04-22)
      Heparan and chondroitin sulfate proteoglycans (HSPGs and CSPGs, respectively) regulate numerous cell surface signaling events, with typically opposite effects on cell function. CSPGs inhibit nerve regeneration through receptor protein tyrosine phosphatase sigma (RPTPσ). Here we report that RPTPσ acts bimodally in sensory neuron extension, mediating CSPG inhibition and HSPG growth promotion. Crystallographic analyses of a shared HSPG-CSPG binding site reveal a conformational plasticity that can accommodate diverse glycosaminoglycans with comparable affinities. Heparan sulfate and analogs induced RPTPσ ectodomain oligomerization in solution, which was inhibited by chondroitin sulfate. RPTPσ and HSPGs colocalize in puncta on sensory neurons in culture, whereas CSPGs occupy the extracellular matrix. These results lead to a model where proteoglycans can exert opposing effects on neuronal extension by competing to control the oligomerization of a common receptor.
    • Proteoglycans in cellular differentiation and neoplasia.

      Gallagher, John T; Hampson, Ian N (1984-06)