• Haemopoietic injury after irradiation: analysis of dose responses and repair using a target-cell model.

      Millar, W T; Hendry, Jolyon H; CRC Department of Radiation Oncology, Glasgow University, UK. (1997-11)
      Published dose-incidence data for haemopoietic lethality in mice have been analysed using a mathematical model based on target-cell survival. The analysis of three comprehensive data sets produced an initial D(o) value of about 1.4 Gy, decreasing to about 1.1 Gy at 3 Gy, 0.9 Gy at 5 Gy, and about 0.7 Gy at a dose of 10 Gy. The alpha/beta ratio was about 14 Gy, and the repair half-time was about 0.3 h. The level of target-cell depletion at LD37 was at about 6 x 10(-4). The D(o) values are compatible with those measured directly for several stages of early haemopoietic progenitor cells in the marrow. The additional use of 13 or alternatively 24 other less-comprehensive data sets increased the overall degree of heterogeneity, so flattening dose-response curves and increasing the deduced overall D(o) values by a factor of about 2. However, when these data sets were stratified with respect to ln(N(o)) where N(o) is the number of tissue rescuing units (TRU), the results were comparable to those obtained when the three comprehensive data sets were analysed individually. Also, the repair halftime was higher at about 1 h. Further, the implied radiosensitivity of the projected target-cell population comprising the TRU was similar to the survival curves obtained for CFU-S and other closely-related haemopoietic progenitor cell types. It has been shown that the number of critical TRU at risk in the marrow is the main feature modulating heterogeneity even when it is assumed that the cellular radiosensitivity does not vary between strains. The number of stem cells comprising a TRU may vary between strains and this may also be influenced by environmental and/or immunological factors. However, it is certainly the case that the initial complement of TRU plays a major role in the incidence of whole body radiation induced mortality.
    • Haemopoietic progenitor and myeloid cell kinetics in humans treated with interleukin-3 and granulocyte/macrophage colony-stimulating factor in combination.

      Lord, Brian I; Testa, Nydia G; Bretti, Sergio; Chang, James; Demuynck, Hilde; Coutinho, Lucia H; De Campos, E S; Fitzsimmons, Lesley; Scarffe, J Howard; Department of Experimental Haematology, PICR, Manchester, UK. (1994-11-15)
      Patients with advanced adenocarcinoma of the colon, rectum or pancreas were entered into trials for evaluation of treatment with sequential doses of IL-3 and GM-CSF. They received 0.25 to 5 micrograms IL-3/kg/d for up to 7 days, followed by 1 microgram GM-CSF/kg/day for a maximum of 10 further days. We assessed the kinetics of bone-marrow cell proliferation and of blood production using tritiated thymidine labelling in vitro and in vivo. Megakaryocytic-CFC were unaffected but proliferation rates of GM-CFC and BFU-E were increased. Progenitor cells were mobilized (12-fold over baseline) into the peripheral blood. The proliferative activity of maturing cells in the marrow was increased (cell-cycle times were reduced by at least 30%). This translated into amplified blood cell production (WCC approximately 30 x 10(9)/l), a 2.2-fold increase in platelet counts and significant eosinophilia. Newly generated neutrophils appeared in the circulation at the normal time and their peripheral half-life was also normal. The calculated 3.2-fold amplification in neutrophil production required nearly 2 extra divisions in the marrow, shared between the progenitors and the proliferating granulocytic cells. The results were compared with those of a previous trial using GM-CSF only, although at a 10-fold higher dose level. Comparable levels of peripheral neutrophils were obtained in both trials but significant ineffective granulopoiesis developed in the earlier study. This was overcome in the present study, the priming dose of IL-3 apparently giving the latitude to utilize lower doses of GM-CSF with less risk of complications.
    • Haemopoietic stem cell development to neutrophils is associated with subcellular redistribution and differential expression of protein kinase C subspecies.

      Shearman, M S; Heyworth, Clare M; Dexter, T Michael; Haefner, B; Owen, P J; Whetton, Anthony D; Department of Biochemistry, Kobe University School of Medicine, Japan. (1993-01)
      Multipotential FDCP-Mix A4 (A4) cells can be induced either to self-renew or to differentiate and develop into mature neutrophils in liquid culture, depending on the haemopoietic growth factors with which they are cultured. When cultured in low concentrations of interleukin 3 (IL-3, 1 unit/ml)) plus Granulocyte Macrophage Colony Stimulating Factor (GM-CSF) and Granulocyte-CSF (G-CSF), A4 cells proliferate with accompanying development to form cells which resemble mature, postmitotic neutrophils. The presence of high concentrations of IL-3 (100 units/ml) blocks the development of A4 cells even in the presence of GM-CSF plus G-CSF. A4 cell development to neutrophils is accompanied by major changes in the expression of protein kinase C (PKC) subspecies in these cells. The predominant subspecies present in multipotent A4 cells, as judged by direct chromatographic analysis, was the type III enzyme (alpha) subspecies, whereas in mature A4 cell neutrophils, the type II (beta I + beta II) enzymes were predominant. Phorbol esters added to immature A4 cells resulted in a proliferative response, but when added to postmitotic A4 cells resembling neutrophils they elicited a large increase in reactive oxygen intermediate production. This suggests that the type III (alpha) subspecies may mediate proliferative responses in stem cells, whilst the type II (beta I + beta II) enzymes are more important for the mature cell functions of postmitotic neutrophils. In cultures containing IL-3 (100 units/ml) both the type III, and also the type II subspecies were predominantly membrane-associated for prolonged periods (> 24 hours).(ABSTRACT TRUNCATED AT 250 WORDS)
    • Haemopoietic stem cell proliferation: spatial and temporal considerations.

      Wright, E; Lord, Brian I; Department of Anatomy and Experimental Pathology, University of St Andrews, Bute Medical Buildings, Westburn Lane, St Andrews, Fife KY16 9TS. (1986)
    • Haemopoietic stem cells

      Lajtha, L G; Paterson Laboratories, Christie Hospital and Holt Radium Institute, Manchester M20 9BX (1975)
    • Haemopoietic stem cells and the problem of self-renewal.

      Dexter, T Michael; Spooncer, Elaine; Schofield, Raymond; Lord, Brian I; Simmons, P J (1984)
      Haemopoiesis occurs in association with a complex stromal cell network in which all levels of haemopoietic cell development can be found. In order to understand the interaction between stromal cells and growth factors with the processes of self-renewal and differentiation, we have carried out a series of experiments attempting to define the circumstances in which self-renewal occurs in long-term marrow cultures. We have found that highly purified (FACS sorted) CFU-S do not undergo significant self-renewal in vitro when inoculated onto marrow stromal cells that can support self-renewal of unfractionated CFU-S. We have examined the effects of expression of the src oncogene on self-renewal of CFU-S. We have found that, following infection of long-term cultures with a retrovirus carrying the src oncogene, there is expression of src in certain of the stromal cells. There is also a selection for CFU-S that have an extended self-renewal capacity in vivo and in vitro. These CFU-S are non-leukaemic and can reconstitute haempoiesis in irradiated mice. Cells from src-infected cultures can also be induced to proliferate and form cell lines in vitro in the presence of interleukin 3 (IL-3). The cell lines produced are multipotential and non-leukaemic. From such data we conclude that expression of the src oncogene has (directly or indirectly) permanently altered the stem cells in such a way that they can undergo extensive self-renewal in situations that are unfavourable for growth and self-renewal of normal stem cells.
    • Haemopoietic Stem-cell Kinetics during Continuous Irradiation.

      Chu-Tse, Wu; Lajtha, L G; Paterson Laboratories, CHristie Hospital and Holt Radium Institute, Manchester (1975)
    • Hair cortical cell counts (HCCC), a new sensitive in vivo assay with possible applications for biological dosimetry.

      Potten, Christopher S; Department of Epithelial Biology, Paterson Institute for Cancer Research, Christie Hospital, (NHS) Trust, Manchester, UK. (1993-01)
      In the region of a growing hair follicle where terminal keratinization takes place, the nuclei of the medulla and cortex are gradually degraded. In the zone between rapid cell proliferation in the matrix and the complete loss of the nuclei in the formed hair, the number of medullary cells in the hair proved to be a sensitive-radiation-dose-dependent endpoint (Potten et al. 1990). Nuclei in the cortex of the hair are similarly degraded but are more difficult to quantitate since they form a circumferential ring of cells round the medulla. However, using the confocal microscope to obtain discrete optical sections just above and below the medulla, the number of cortical cells was reproducibly counted. The maximum reduction in cortical cell count occurred on the 3rd day after irradiation. Here, the dose-response curve (0.5-4.0 Gy) is presented, for the population of cortical cells in awl guard hairs of the mouse for 137Cs gamma-rays at 4 Gy/min, which was exponential with no significant shoulder, (the D0 was 2.4 +/- 0.2 Gy and the extrapolation number 1.2 +/- 0.1). The cortical cells exhibited a slightly higher D0 value than the medullary cells but differed in that there was no evidence of a shoulder. The effects of 0.5 Gy could be detected. The degree of sensitivity, and the fact that cortical cells are readily detected in human hair follicles, unlike the medullary cells, make this a potentially valuable human biological dosimeter.
    • Hair medullary cell counts following low-dose-rate gamma- and high-energy neutron irradiation.

      Potten, Christopher S; Cancer Research Campaign Department of Epithelial Biology, Paterson Institute for Cancer Research, Christie Hospital, (NHS) Trust, Manchester, UK. (1993-01)
      Young adult Balb/c mice with hair follicles synchronously in the middle of the hair growth cycle received whole-body or partial-body doses of gamma-radiation or neutron radiation. The hair follicles were analysed either 3 days after irradiation in the dose-response experiments, or at various times after a constant dose in the time-course experiments, for changes in the number of cells in the forming medulla of the hair in the region just above the germinal matrix of the growing (anagen) hair follicle. Time-course experiments showed that 3 days after irradiating growing follicles (2 or 4 Gy of gamma-rays or 1 or 2 Gy of neutrons), the maximum reduction in the hair medullary cell count (HMCC) was observed. Survival curves were obtained for gamma-rays over a range of dose-rates (4.0-0.0023 Gy/min) using total doses between 0.5 and 5.0 Gy. A survival curve was also obtained for 62 MeV neutrons at a dose-rate of 0.31 Gy/min and doses of 0.1-2.0 Gy. The D0 for the HMCC dose-response curve following caesium gamma-irradiation at 4.0 Gy/min was 2.1 +/- 0.2 Gy with n = 1.7 +/- 0.2. The dose-response curve for low-dose-rate gamma-rays was best fitted by a simple exponential function with no evidence of a shoulder. The D0 was 3.0 +/- 0.1 Gy at a dose-rate of 0.04 Gy/min. The data for 62 MeV neutrons (at 0.35 Gy/min) had a small but significant shoulder n = 1.5 +/- 0.1 and a D0 of 1.0 +/- 0.1 Gy. These data further illustrate the sensitivity of this assay and its potential application as a biological dosimeter.
    • Hair medullary cell counts: a simple and sensitive indicator of radiation exposure.

      Potten, Christopher S; Geng, L; Taylor, P; Paterson Institute for Cancer Research, Christie Hospital, Manchester, U.K. (1990-01)
      The medulla in the lower regions of a growing mouse hair contains a very regularly spaced column of cell nuclei. The total number of nuclei 3 days after irradiation in this column (from its lower recognition point to the point of terminal differentiation and nuclear degradation) proves to be a sensitive indicator of the level of radiation exposure.
    • HALT: targeted therapy with or without dose-intensified radiotherapy in oligo-progressive disease in oncogene addicted lung tumours.

      McDonald, F; Guckenberger, M; Popat, S; Faivre-Finn, Corinne; Andratschke, N; Riddell, A; Hanna, G; Franks, K; Miles, E; Patel, R; et al. (2018-01)
    • Haploinsufficiency of Runx1 results in the acceleration of mesodermal development and hemangioblast specification upon in vitro differentiation of ES cells.

      Lacaud, Georges; Kouskoff, Valerie; Trumble, Anne; Schwantz, Staci; Keller, Gordon; Carl C. Icahn Center for Gene Therapy and Molecular Medicine, Mount Sinai School of Medicine, New York, NY 10029, USA. (2004-02-01)
      The AML1 gene (recently renamed Runx1), which encodes the DNA-binding subunit of a transcription factor of the core binding factor (CBF) family, is required for the establishment of definitive hematopoiesis. We have previously demonstrated that Runx1 is expressed in yolk sac mesodermal cells prior to the establishment of the blood islands and in the embryoid body (EB)-derived blast-colony-forming cells (BL-CFCs), the in vitro equivalent of the hemangioblast. Analysis of Runx1-deficient embryonic stem (ES) cells demonstrated that this gene is essential for the generation of normal numbers of blast colonies, the progeny of the BL-CFCs. In the present study, we analyzed the potential of Runx1(+/-) ES cells to determine if heterozygosity at the Runx1 locus impacts early developmental events leading to the commitment of the BL-CFCs. Our results indicate that Runx1 heterozygosity leads to an acceleration of mesodermal commitment and specification to the BL-CFCs and to the hematopoietic lineages in EBs.
    • Harmonization of programmed cell death ligand-1 diagnostic assays in non-small cell lung cancer

      Popple, Amy; Illidge, Timothy M; Targeted Therapy Group, Division of Molecular and Clinical Cancer Sciences, The University of Manchester, Manchester Cancer Research Centre, Manchester. UK (2017-05)
    • Harnessing immunity for therapy in human papillomavirus driven cancers

      Stern, Peter L; Manchester Cancer Research Centre, University of Manchester, Manchester, M20 4GJ (2021)
      In persistent high-risk HPV infection, viral gene expression can trigger some important early changes to immune capabilities which act to protect the lesion from immune attack and subsequently promote its growth and ability for sustained immune escape. This includes immune checkpoint-inhibitor ligand expression (e.g. PD-L1) by tumour or associated immune cells that can block any anti-tumour T-cell effectors. While there are encouraging signs of efficacy for cancer immunotherapies including with immune checkpoint inhibitors, therapeutic vaccines and adoptive cell therapies, overall response and survival rates remain relatively low. HPV oncogene vaccination has shown some useful efficacy in treatment of patients with high-grade lesions but was unable to control later stage cancers. To maximally exploit anti-tumour immune responses, the suppressive factors associated with HPV carcinogenesis must be countered. Importantly, a combination of chemotherapy, reducing immunosuppressive myeloid cells, with therapeutic HPV vaccination significantly improves impact on cancer treatment. Many clinical trials are investigating checkpoint inhibitor treatments in HPV associated cancers but response rates are limited; combination with vaccination is being tested. Further investigation of how chemo- and/or radio-therapy can influence the recovery of effective anti-tumour immunity is warranted. Understanding how to optimally deploy and sequence conventional and immunotherapies is the challenge.
    • HBP2: a new mammalian protein that complements the fission yeast MBF transcription complex.

      Sánchez-Díaz, Alberto; Blanco, Miguel A; Jones, Nic; Moreno, Sergio; Instituto de Microbiologia Bioquimica, Departamento de Microbiologia y Genética, CSIC/Universidad de Salamanca, Spain. (2001-09)
      The mammalian HBP2 gene has been isolated by functional complementation of cells unable to undergo DNA replication in fission yeast. HBP2 is a protein with a high mobility group (HMG) domain that belongs to the Sox (Sry-related HMG box) family of transcription factors. As in other members of the family, the HMG box in HBP2 is a DNA-binding domain that is essential for its function in Schizosaccharomyces pombe. Expression of HBP2 in fission yeast activates CdclO-dependent transcription at G1/S and allows growth of cdc10-129, resldelta and rep2delta mutant cells at the restrictive temperature. The mammalian HBP2 activates transcription at G1/S in the fission yeast Sch. pombe.
    • HDAC1 and HDAC2 modulate TGF-β signaling during endothelial-to-hematopoietic transition.

      Thambyrajah, Roshana; Fadlullah, Muhammad Z H; Proffitt, Martin; Patel, Rahima; Cowley, S; Kouskoff, Valerie; Lacaud, Georges; CRUK Stem Cell Biology Group, CRUK Manchester Institute, 555 Wilmslow Road, Manchester M20 4GJ, UK (2018-04-10)
      The first hematopoietic stem and progenitor cells are generated during development from hemogenic endothelium (HE) through trans-differentiation. The molecular mechanisms underlying this endothelial-to-hematopoietic transition (EHT) remain poorly understood. Here, we explored the role of the epigenetic regulators HDAC1 and HDAC2 in the emergence of these first blood cells in vitro and in vivo. Loss of either of these epigenetic silencers through conditional genetic deletion reduced hematopoietic transition from HE, while combined deletion was incompatible with blood generation. We investigated the molecular basis of HDAC1 and HDAC2 requirement and identified TGF-β signaling as one of the pathways controlled by HDAC1 and HDAC2. Accordingly, we experimentally demonstrated that activation of this pathway in HE cells reinforces hematopoietic development. Altogether, our results establish that HDAC1 and HDAC2 modulate TGF-β signaling and suggest that stimulation of this pathway in HE cells would be beneficial for production of hematopoietic cells for regenerative therapies.
    • Head scatter modelling for irregular field shaping and beam intensity modulation.

      Hounsell, Alan R; Wilkinson, John M; North Western Medical Physics, Christie Hospital NHS Trust, Manchester, UK. phyarh@picr.cr.man.ac.uk (1997-09)
      Scattered radiation from within the treatment head can contribute significant dose to all parts of a radiotherapy treatment field. A multileaf collimator may be used to create an arbitrarily shaped field, and may also be used, under dynamic control, to modulate the beam intensity over the field. This method of intensity modulation is effectively a superposition of a large number of fields which have the same beam direction, but different shapes, and some of these shapes may have unusually small dimensions, particularly in the direction of the leaf movement. Two models for predicting the head scatter under these conditions have been investigated. These are a first-order Compton scatter approximation from the flattening filter, and an empirical fit to measured data using an exponential function. The first model only considers scatter from the flattening filter and has been applied to field sizes between 2 cm by 2 cm and 10 cm by 10 cm, where agreements are all within 1%. However it is not satisfactory at larger field sizes where small scatter contributions, from scattering sources other than the flattening filter, are integrated over large areas. The second model uses measured data between 4 cm by 4 cm and 30 cm by 30 cm to optimize the exponential function and is used to calculate the head scatter contribution for all field sizes. In this case good agreement is achieved over the full field size range, and hence this is a more generally applicable model. Results are presented for static irregularly shaped fields and intensity modulated beams created using a Philips multileaf collimator.
    • Health-related quality of life and work productivity in UK patients with HER2-positive breast cancer: a cross-sectional study evaluating the relationships between disease and treatment stage

      Verrill, M.; Wardley, Andrew M; Retzler, J.; Smith, A. B.; Bottomley, C.; S, N. D.; Tran, I.; Leslie, I.; Schmid, P.; The Newcastle Upon Tyne Hospitals NHS Foundation Trust, Newcastle, UK. (2020)
      Background: The impact of different disease stages and treatment for human epidermal growth factor 2 positive (HER2-positive) breast cancer (BC) on work productivity and health-related quality of life (HRQoL) is poorly understood. Methods: This was a UK cross-sectional study of 299 adult patients with HER2-positive early or metastatic BC (NCT03099200). Productivity was assessed using the work productivity and activity impairment scale; HRQoL was measured using EuroQol-5 Dimensions-5 levels (EQ-5D-5L), and Functional Assessment of Cancer Therapy Breast (FACT-G and -B) instruments. Three balanced patient groups were recruited: (1) early BC on treatment post-surgery, (2) early BC after completion of adjuvant treatment, (3) during metastatic BC treatment. Between-group comparisons were performed using an analysis of variance. Results: Group 1 comprised 89 patients, Group 2, 108 and Group 3, 102. Age, ethnicity and comorbidities were similar across groups. Patients in Group 3 reported more often being unable to work (significant Bonferroni adjusted p < 0.003). Proportions of employed patients were 50.6%, 50.9% and 27.5% in Groups 1, 2 and 3, respectively. For patients in part-time employment, the number of hours worked was significantly higher in Group 2 patients versus Group 3 (p = 0.002). Group 2 also had significantly lower levels of work absenteeism and overall work impairment compared with Group 1 (p < 0.001). Patients in Group 3 reported worse health utility scores (p ≤ 0.002), moderate or worse problems in the EQ-5D-5L self-care and usual activity domains (p ≤ 0.001), and lower HRQoL as assessed by FACT summary scores (p < 0.001 for FACT-B and -G) than Groups 1 and 2. Poorer HRQoL was significantly associated with higher work impairment (p < 0.001), with the strongest relationships being observed between activity impairment and HRQoL (Pearson's r: 0.67). Conclusions: Metastatic disease and treatment of HER2-positive BC adversely impacted on work productivity and HRQoL. The results of this study support the idea that being able to delay or prevent the metastatic recurrence of BC, for example by extending the time patients are in remission or at early stage of BC, has wider benefits in terms of patient productivity and HRQoL.
    • Heart position reproducibility in deep inspiration breath hold radiotherapy for lung cancer

      Josipovic, M; Aznar, Marianne Camille; Thomsen, JB; Scherman, J; Damkjaer, SMS; Nygard, L; Specht, L; Pohl, M; Persson, GF; Rigshospitalet, Dept. of Oncology\Section of Radiotherapy, Copenhagen, Denmark (2019)