Nicol Keith

University of Glasgow | UofG · Institute of Cancer Sciences

The Sustainable Development and the Global South project was jointly conceived by the Innovation School at Glasgow School of Art in partnership with the Sustainable Futures in Africa Network (SFA), and the University of Glasgow. This project asked students to consider what happens in this global landscape ten years from now where Sustainable Devel...

The Precision Medicine and the Future of Cancer project was jointly conceived by the Innovation School at Glasgow School of Art and the Institute of Cancer Sciences at the University of Glasgow. This project asked students to consider what will happen in a cancer landscape ten years from now, where PM has evolved to the extent that new forms of me...

The FUTURE of CANCER and COLLECTIVE INTELLIGENCE in the POST-COVID WORLD project is a highly innovative and creative student-led project. The Glasgow School of Art Innovation School’s final year Product Design students and faculty formed a dynamic community of practice with cancer practitioners and researchers from the University of Glasgow & beyo...

The COVID-19 pandemic has brought into focus the complexity and interrelatedness of the range of health care delivery associated with translational cancer research. Importantly it highlights the potential fragility and also strengths of this ecosystem. The ECMC North Education Event was an online education event, held on the 19th of October 2020,...

The Sustainable Development Goals, as defined by the United Nations’ 2030 Agenda for Sustainable Development, provide recognised framework for wider impact, allowing research institutions worldwide to demonstrate the relevance of their output to societal challenges. Using the SDG to show commitment to sustainable development and innovation in an in...

Bringing new cancer therapies into routine cancer care is a long and complex process. Phase 1 clinical trials assess new cancer treatments in people for the first time. It is a critical step in the pathway. The aim of this project is to make the world of a cancer Phase 1 trialist and the value they bring, understandable to others. From those intere...

Telomere signaling and metabolic dysfunction are hallmarks of cell aging. New agents targeting these processes might provide therapeutic opportunities, including chemoprevention strategies against cancer predisposition. We report identification and characterization of a pyrazolopyrimidine compound series identified from screens focused on cell immo...

Accomplissh (‘Accelerate co-creation by setting up a multi-actor platform for impact from social sciences and humanities’) is a unique co-creation engagement platform focused on societal, cultural, economic or policy-related impacts originating from social sciences and humanities research. It is funded through the EU Horizon 2020 research and innov...

Senescence is a universal barrier to immortalisation and tumorigenesis. As such, interest in the use of senescence-induction in a therapeutic context has been gaining momentum in the past few years; however, senescence and immortalisation remain underserved areas for drug discovery owing to a lack of robust senescence inducing agents and an incompl...

Expression of senescence biomarkers allows validation and refinement of screen hits. Validation of the top 40 hit siRNAs for nuclear area increase in A375P (a) and HCT116 (b) ranked in descending order. Mean nuclear area per well (μm2) for triplicate wells in 3 independent transfections is represented as box whisker plots generated in Tableau deskt...

Set-up of a morphology based screen for siRNAs inducing a senescent phenotype. (a) Screen overview. A375P melanoma cells were transfected with 50nM siRNA from the Ambion Druggable Genome Library. 5 days later cells were fixed, stained for DAPI and imaged using the Operetta high content imaging platform. Nuclei in acquired images were detected and q...

A large-scale morphology screen identifies siRNAs that induce a senescent-like morphology in A375P melanoma cells. (a) Scatterplot showing results of a rescreen of 810 siRNAs representing all 3 siRNA oligos included in the Ambion druggable genome library for the top 270 gene hits in A375P. (b) Scatterplot showing the position of all 3 siRNAs target...

Expression of p21 and 53BP1 associated with senescence inducing siRNAs. Representative images of p21 (top) and 53BP1 (bottom) staining for siRNA hits and scrambled controls in A375P (a) and HCT116 (b). (TIF)

Supporting methods, figure legends, and tables. Supporting methods include details of cell lines and culture methods, siRNA transfection methods, screen set-up, statistical analyses and antibodies. Supporting table A shows Operetta analysis sequences and algorithms used. Supporting table B shows parameters used for ArrayScan analysis. (DOCX)

Caspase 3/7 activity and ECT2 levels after ECT2 knockdown in KRAS mutant HCT116 parental cells. (a) Promega Apo-ONE caspase 3/7 assay in HCT116 parental cells. Cells were left untransfected (cells) or were transfected with ECT2, CDK1, or scrambled siRNA. Cell culture medium was included as negative control. Assays were performed 5 days post-transfe...

Validation of the cellular senescence screen. This file provides details of assays performed using etoposide and a kinase inhibitor library to determine optimal phenotypic parameters for use in the screen. Senescence was initially evaluated in A375P cells by a range of assays including growth, colony formation, SAβGal, p21 and 53BP1, H2AX and nucle...

Barrett’s oesophagus is a premalignant metaplastic condition that predisposes patients to the development of oesophageal adenocarcinoma. However, only a minor fraction of Barrett’s oesophagus patients progress to adenocarcinoma and it is thus essential to determine bio-molecular markers that can predict the progression of this condition. Telomere d...

Cancer immune evasion is a major stumbling block in designing effective anticancer therapeutic strategies. Although considerable progress has been made in understanding how cancers evade destructive immunity, measures to counteract tumor escape have not kept pace. There are a number of factors that contribute to tumor persistence despite having a n...

Apoptosis or programmed cell death is natural way of removing aged cells from the body. Most of the anti-cancer therapies trigger apoptosis induction and related cell death networks to eliminate malignant cells. However, in cancer, de-regulated apoptotic signaling, particularly the activation of an anti-apoptotic systems, allows cancer cells to esc...

Targeted therapies and the consequent adoption of “personalized” oncology have achieved notable successes in some cancers; however, significant problems remain with this approach. Many targeted therapies are highly toxic, costs are extremely high, and most patients experience relapse after a few disease-free months. Relapses arise from genetic hete...

Genomic instability can initiate cancer, augment progression, and influence the overall prognosis of the affected patient. Genomic instability arises from many different pathways, such as telomere damage, centrosome amplification, epigenetic modifications, and DNA damage from endogenous and exogenous sources, and can be perpetuating, or limiting, t...

Cellular senescence is a barrier to tumorigenesis in normal cells, and tumor cells undergo senescence responses to genotoxic stimuli, which is a potential target phenotype for cancer therapy. However, in this setting, mixed-mode responses are common with apoptosis the dominant effect. Hence, more selective senescence inducers are required. Here we...

Cellular senescence is a widespread stress response and is widely considered to be an alternative cancer therapeutic goal. Unlike apoptosis, senescence is composed of a diverse set of subphenotypes; depending on which of its associated effector programs are engaged. Here we establish a simple and sensitive, cell-based, prosenescence screen with det...

One of the hallmarks of malignant cell populations is the ability to undergo continuous proliferation. This property allows clonal lineages to acquire sequential aberrations that can fuel increasingly autonomous growth, invasiveness, and therapeutic resistance. Innate cellular mechanisms have evolved to regulate replicative potential as a hedge aga...

Cancer is a key health issue across the world, causing substantial patient morbidity and mortality. Patient prognosis is tightly linked with metastatic dissemination of the disease to distant sites, with metastatic diseases accounting for a vast percentage of cancer patient mortality. While advances in this area have been made, the process of cance...

The evasion of anti-growth signaling is an important characteristic of cancer cells. In order to continue to proliferate, cancer cells must somehow uncouple themselves from the many signals that exist to slow down cell growth. Here, we define the anti-growth signaling process, and review several important pathways involved in growth signaling: p53,...

Deregulation of angiogenesis–the growth of new blood vessels from an existing vasculature–is a main driving force in many severe human diseases including cancer. As such, tumor angiogenesis is important for delivering oxygen and nutrients to growing tumors, and therefore considered an essential pathologic feature of cancer, while also playing a key...

Background: N-acetyl-glucosaminidase (NAG) is a potential marker of genotoxicity. We retrospectively analyzed plasma NAG and clinico-pathologic features in advanced gastrointestinal adenocarcinoma patients. Methods: Plasma from 118 patients and 51 healthy volunteers was analyzed for associations between NAG levels and age, disease presence, stag...

We have previously described gene expression changes during spontaneous immortalization of T-cells, thereby identifying cellular processes important for cell growth crisis escape and unlimited proliferation. Here, we analyze the same model to investigate the role of genome-wide methylation in the immortalization process at different time points pre...

Located in the basal epidermis and hair follicles, melanocytes of the integument are responsible for its coloration through production of melanin pigments. Melanin is produced in lysosomal-like organelles called melanosomes. In humans, this skin pigmentation acts as an ultraviolet radiation filter. Abnormalities in the division of melanocytes are q...

Cancer cells depend on transcription of telomerase reverse transcriptase (TERT). Many transcription factors affect TERT, though regulation occurs in context of a broader network. Network effects on telomerase regulation have not been investigated, though deeper understanding of TERT transcription requires a systems view. However, control over indiv...

Cellular senescence has attracted renewed interest in recent years in light of the realization that cancer cells are susceptible to senescence-like responses to stress including exposure to chemotherapeutic agents. Therefore, senescence is viewed as a potentially important target in cancer therapy. However, translation of senescence therapeutics wi...

Human mesenchymal stem cells (hMSCs) have been shown to have potential in regenerative approaches in bone and blood. Most protocols rely on their in vitro expansion prior to clinical use. However, several groups including our own have shown that hMSCs lose proliferation and differentiation ability with serial passage in culture, limiting their clin...

This study investigated the prevalence and the prognostic relevance of the 2 known telomere maintenance mechanisms (TMMs), telomerase activity (TA) and alternative lengthening of telomeres (ALT), in malignant peripheral nerve sheath tumors (MPNST). In 57 specimens from 49 patients with MPNST (35 sporadic, 14 neurofibromatosis type 1-related), TA wa...

The field of stem cell biology continues to evolve by characterization of further types of stem cells and by exploring their therapeutic potential for experimental and clinical applications. Human mesenchymal stem cells (hMSCs) are one of the most promising candidates simply because of their easiness of both ex vivo expansion in culture dishes and...

Oncogene is one of the world’s leading cancer journals. It is published weekly and covers all aspects of the structure and function of Oncogenes.

Stem cells are responsible for tissue repair and maintenance and it is assumed that changes observed in the stem cell compartment with age underlie the concomitant decline in tissue function. Studies in murine models have highlighted the importance of intrinsic changes occurring in stem cells with age. They have also drawn the attention to other fa...

Cell senescence is a permanent growth arrest following extended proliferation. Cultured cancer cells including metastatic melanoma cells often appear immortal (proliferate indefinitely), while uncultured benign nevi (moles) show senescence markers. Here, with new explantation methods, we investigated which classes of primary pigmented lesions are t...

Aberrant telomere homeostasis is essential for cell immortality, enabling cells to evade telomere dependent senescence. Disruption of telomere structure and function in cancer cells is highly toxic as shown by detailed pre-clinical evaluation of telomerase inhibitors. Under telomerase inhibition, cells must divide sufficiently frequently to allow o...

One of the fundamental changes required for tumorigenesis is escape from cellular senescence. Strategies to induce senescence in cancer cells might provide future therapies complementary to existing interventions aimed at apoptosis. Progress toward senescence targeted drug discovery could be accelerated by applying novel screening-technologies and...

Cellular senescence is a major barrier to tumour progression, though its role in pathogenesis of cancer and other diseases is poorly understood in vivo. Improved understanding of the degree to which latent senescence signalling persists in tumours might identify intervention strategies to provoke "accelerated senescence" responses as a therapeutic...

Artificial transcription factors (ATFs) consist of a transcriptional effector domain fused to a DNA-binding domain such as an engineered zinc finger protein (ZFP). Depending on the effector domain, ATFs can up- or downregulate gene expression and thus represent powerful tools in biomedical research and allow novel approaches in clinical practice. H...

Replicative senescence forms a major barrier to tumor progression. Cancer cells bypass this by using one of the two known telomere maintenance mechanisms: telomerase or the recombination-based alternative lengthening of telomeres (ALT) mechanism. The molecular details of ALT are currently poorly understood. We have previously shown that telomerase...

Our knowledge on immortalization and telomere biology is mainly based on genetically manipulated cells analyzed before and many population doublings post growth crisis. The general view is that growth crisis is telomere length (TL) dependent and that escape from crisis is coupled to increased expression of the telomerase reverse transcriptase (hTER...

Telomere length is maintained by two known mechanisms, the activation of telomerase or alternative lengthening of telomeres (ALT). The molecular mechanisms regulating the ALT phenotype are poorly understood and it is unknown how the decision of which pathway to activate is made at the cellular level. We have shown earlier that active repression of...

DJ-1 is as a novel regulator of the tumor suppressor PTEN with stimulatory effects on PI3K-AKT/PKB signaling, one possible target of which is cMyc. The catalytic unit of the telomerase complex, hTERT, can be activated at different levels, including transcriptionally by cMyc and through phosphorylation by AKT/PKB. The aim of the study was to analyze...

Chemical structures of GSK3 inhibitors reported in the study. (0.14 MB TIF)

Regulation of Topflash reporter activity by over-expression of Wnt pathway components. (0.15 MB TIF)

Optimisation of the best-fit transcriptional network. (1.23 MB TIF)

BIO selectively represses expression of the full length hTERT transcript in A2780. (0.94 MB TIF)

Differentially expressed Agilent IDs and fold intensity change. (0.09 MB XLS)

Legends to supporting figures and files (0.03 MB DOC)

BIO, but not MeBIO, activates β-catenin signalling. (0.07 MB TIF)

Expression of hTERT after 16 h treatment with 5 µM BIO (1.92 MB TIF)

Representative results of MetaCore “transcriptional-regulation” algorithm analysis. (3.60 MB TIF)

Antibodies used in the study. (0.02 MB XLS)

Best fit transcriptional network statistics. (0.21 MB XLS)

Multiple mechanisms of senescence induction exist including telomere attrition, oxidative stress, oncogene expression and DNA damage signalling. The regulation of the cellular changes required to respond to these stimuli and create the complex senescent cell phenotype has many different mechanisms. MiRNAs present one mechanism by which genes with d...

Stem cells are central to the development and maintenance of many tissues. This is due to their capacity for extensive proliferation and differentiation into effector cells. More recently it has been shown that the proliferative and differentiative ability of stem cells decreases with age, suggesting that this may play a role in tissue aging. Down...

In this study, we show that NANOG, a master transcription factor, regulates S-phase entry in human embryonic stem cells (hESCs) via transcriptional regulation of cell cycle regulatory components. Chromatin immunoprecipitation combined with reporter-based transfection assays show that the C-terminal region of NANOG binds to the regulatory regions of...

Telomerase controls telomere homeostasis and cell immortality and is a promising anti-cancer target, but few small molecule telomerase inhibitors have been developed. Reactivated transcription of the catalytic subunit hTERT in cancer cells controls telomerase expression. Better understanding of upstream pathways is critical for effective anti-telom...

Marking of human embryonic stem (ES) and embryonal carcinoma (EC) cells with pluripotent promoter-driven reporter gene cassettes provides an important tool for studies related to maintenance of pluripotency, cell differentiation and cell selection. OCT4, TERF1 and telomerase reverse transcriptase component (TERT) are considered as pluripotent marke...