Jenny Southgate

The University of York · Department of Biology

Interferon gamma (IFNγ) is central to the inflammatory immune response, such as that entrained by BCG immunotherapy for bladder cancer. However, immune-mediated tumour cell killing is subject to modulation by immunoinhibitory “checkpoint” receptors such as PD-L1. We investigated the effects of IFNγ on barrier-forming in vitro-differentiated normal...

Urothelium is a transitional, stratified epithelium that lines the lower urinary tract, providing a tight barrier to urine whilst retaining the capacity to stretch and rapidly resolve damage. The role of glycerophospholipids in urothelial barrier function is largely unknown, despite their importance in membrane structural integrity, protein complex...

Limited understanding of bladder cancer aetiopathology hampers progress in reducing incidence. Mutational signatures show the anti-viral apolipoprotein B mRNA editing enzyme catalytic polypeptide (APOBEC) enzymes are responsible for the preponderance of mutations in bladder tumour genomes, but no causative viral agent has been identified. BK polyom...

Bladder exstrophy (BEX) is a rare developmental abnormality resulting in an open exposed bladder plate. Whilst normal bladder urothelium is a mitotically-quiescent barrier epithelium, histological studies of BEX epithelia report squamous and proliferative changes that can persist beyond surgical closure. We examined whether patient-derived BEX epit...

Intravesical Bacillus Calmette-Guérin vaccine (BCG) is an established immunotherapeutic in bladder cancer (BlCa), provoking inflammation leading to tumour-specific immunity. Immune checkpoint blockers such as anti-PD-L1 have potential for enhancing tumour-specific lymphocyte-mediated cytotoxicity in BCG-refractive or advanced disease. In both cases...

Bladder acellular matrix has promising applications in urological and other reconstructive surgery as it represents a naturally compliant, non-immunogenic and highly tissue-integrative material. As the bladder fills and distends, the loosely-coiled bundles of collagen fibres in the wall become extended and orientate parallel to the lumen, resulting...

Limited understanding of bladder cancer aetiopathology hampers progress in reducing incidence. BK polyomavirus (BKPyV) is a common childhood infection that can be reactivated in the adult kidney leading to viruria. Here we used a mitotically-quiescent, differentiated, normal human urothelial in vitro model to study BKPyV infection. BKPyV infection...

Acellular matrices produced by tissue decellularisation are reported to have tissue integrative properties. We examined the potential for incorporating acellular matrix grafts during procedures where there is an inadequate natural tissue bed to support an enduring surgical repair. Hypospadias is a common congenital defect requiring surgery, but ass...

This chapter reviews the use of engineering and biomaterials approaches aimed at improving the capacity and compliance of the diseased urinary bladder. The chapter introduces the normal bladder and describes the clinical background that drives the need for bladder tissue replacement and/or reconstruction. It examines a variety of synthetic and natu...

Introduction Smoking is the best established risk factor for bladder cancer and direct mutagenesis by smoke carcinogens on the cellular genome can be described in clonal populations as mutational signatures. A mutational signature summarises a lifetime of DNA-damage (caused by interaction between mutagenic processes, tissue-specific gene transcript...

Disparity between genome-wide mutations in bladder and other cancers where smoking is a risk factor raises questions about carcinogenesis in different epithelia. To develop an experimental model of bladder carcinogenesis, we clonally expanded in vitro differentiated normal human urothelial (NHU) cells following exposure to an exemplar procarcinogen...

Inherited Primary Immunodeficiency (PID) disorders are associated with increased risk of malignancy that may relate to impaired antitumor immune responses or a direct role for PID germline mutations in tumorigenesis. We recently identified germline loss of function mutations in Janus Associated Kinase 1 (JAK1) causing primary immunodeficiency chara...

Occupational and environmental exposure to cadmium is associated with the development of urothelial cancer. The metallothionein (MT) family of genes encodes proteins that sequester metal ions and modulate physiological processes, including zinc homeostasis. Little is known about the selectivity of expression of the different MT isoforms. Here, we e...

Introduction: Urothelial cells harvested from benign diseased bladders have a compromised capacity to propagate or differentiate in vitro, potentially limiting their application in autologous tissue engineering approaches. The causative pathways behind this altered phenotype are unknown. The hypothesis is that hypoxic damage to the urothelium occu...

Table S1. Clinical characteristics and histological assessment of MIBC specimens included in the study.

Figure S1. Full western blots for Figure 4A&B. Figure S2. Full western blots for Figure 4A&C. Figure S3. Representative images of immunoperoxidase labelling for AHR and the urothelial barrier proteins, claudin 5 and uroplakin 3a. Figure S4. Differential regulation of CYP1A1 and CYP1B1 transcripts by AHR stimulation of NHU cells in different stat...

Identification of transcription factors expressed by differentiated cells is informative not only of tissue-specific pathways, but to help identify master regulators for cellular reprogramming. If applied, such an approach could generate healthy autologous tissue-specific cells for clinical use where cells from the homologous tissue are unavailable...

Telomerase activity imparts eukaryotic cells with unlimited proliferation capacity, one of the cancer hallmarks. Over 90% of human urothelial carcinoma of the bladder (UCB) tumours are positive for telomerase activity. Telomerase activation can occur through several mechanisms. Mutations in the core promoter region of the human telomerase reverse t...

Open Access Full Text available (onlinelibrary.wiley.com/doi/10.1002/mc.22784/abstract). Extra-hepatic metabolism of xenobiotics by epithelial tissues has evolved as a self-defence mechanism but has potential to contribute to the local activation of carcinogens. Bladder epithelium (urothelium) is bathed in excreted urinary toxicants and pro-carci...

A correction to this article has been published and is linked from the HTML version of this paper. The error has been fixed in the paper.

Vesicoureteric reflux (VUR) is the commonest urological anomaly in children. Despite treatment improvements, associated renal lesions - congenital dysplasia, acquired scarring or both - are a common cause of childhood hypertension and renal failure. Primary VUR is familial, with transmission rate and sibling risk both approaching 50%, and appears h...

The identification of the host defence peptides as target effectors in the innate defence of the uro-genital tract creates new translational possibilities for immunomodulatory therapies, specifically vaginal therapies to treat women suffering from rUTI, particularly those carrying the TLR5_C1174T SNP. Urinary tract infections (UTIs) are a microbial...

Cell differentiation is affected by complex networks of transcription factors that co-ordinate re-organisation of the chromatin landscape. The hierarchies of these relationships can be difficult to dissect. During in vitro differentiation of normal human uro-epithelial cells, formaldehyde-assisted isolation of regulatory elements (FAIRE-seq) and RN...

Telomerase activity imparts eukaryotic cells with unlimited proliferation capacity, one of the cancer hallmarks. Over 90% of human urothelial carcinoma of the bladder (UCB) are positive for telomerase activity. Telomerase activation can occur through several mechanisms. Mutations in the core promoter region of the human telomerase reverse transcrip...

CD40, a member of the tumour necrosis factor receptor (TNFR) superfamily, has the capacity to cause extensive apoptosis in carcinoma cells, while sparing normal epithelial cells. Yet, apoptosis is only achieved by membrane-presented CD40 ligand (mCD40L), as soluble receptor agonists are but weakly pro-apoptotic. Here, for the first time we have ide...

This paper presents a novel method for tracking and characterizing adherent cells in monolayer culture. A system of cell tracking employing computer vision techniques was applied to time-lapse videos of replicate normal human uro-epithelial cell cultures exposed to different concentrations of adenosine triphosphate (ATP) and a selective purinergic...

Recreational abuse of ketamine has been associated with the emergence of a new bladder pain syndrome, ketamine-induced cystitis, characterized by chronic inflammation and urothelial ulceration. We investigated the direct effects of ketamine on normal human urothelium maintained in organ culture or as finite cell lines in vitro. Exposure of urotheli...

Objective: To establish whether the urothelial ulceration observed in ketamine-induced cystitis is triggered by urinary or systemic factors. This was achieved with a rare case where an urachal cyst was found near the bladder dome in a patient undergoing cystectomy for unremitting pain following ketamine abuse. Methods: Clinical investigations in...

This paper presents a novel method for tracking and characterizing adherent cells in monolayer culture. A system of cell tracking employing computer vision techniques was applied to time-lapse videos of replicate normal human uro-epithelial cell cultures exposed to different concentrations of adenosine triphosphate (ATP), acquired over a 20 h perio...

Epithelial tissue structure is the emergent outcome of the interactions between large numbers of individual cells. Experimental cell biology offers an important tool to unravel these complex interactions, but current methods of analysis tend to be limited to mean field approaches or representation by selected subsets of cells. This may result in bi...

Objective: Tight junctions are multicomponent structures, with claudin proteins defining paracellular permeability. Claudin 3 is a candidate for the exceptional "tightness" of human urothelium, being localised to the terminal tight junction (TJ) of superficial cells. Our aim was to determine whether claudin 3 plays an instigating and/or a function...

Summary Extracting relevant information from large-scale data offers unprecedented opportunities in cancerology. We applied independent component analysis (ICA) to bladder cancer transcriptome data sets and interpreted the components using gene enrichment analysis and tumor-associated molecular, clinicopathological, and processing information. We i...

The invention provides an improved method of producing a natural, acellular matrix scaffold for subsequent use in tissue-engineered replacement of tissues such as the bladder. Decellularization is carried out on an expanded or distended bladder and the product retains the strength and compliance of natural material. The invention also provides use...

Muscle-invasive bladder carcinoma (MIBC) constitutes a heterogeneous group of tumors with a poor outcome. Molecular stratification of MIBC may identify clinically relevant tumor subgroups and help to provide effective targeted therapies. From seven series of large-scale transcriptomic data (383 tumors), we identified an MIBC subgroup accounting for...

Regenerative medicine is an emerging field that is focused on the repair, replacement or regeneration of tissues and organs. It involves multiple disciplines dedicated to delivering different aspects of the regeneration process, including cell biology, material sciences and bioengineering. The development of tissue engineering strategies incorporat...

By operating as both a subunit of the cadherin complex and a key component of Wnt signalling, β-catenin constitutes the lynchpin between cell:cell contact and transcriptional regulation of proliferation to co-ordinate epithelial tissue homeostasis and regeneration. Integration of multiple growth-regulatory inputs with β-catenin signalling has been...

The bladder is an important tissue in which to evaluate xenobiotic drug interactions and toxicities due to the concentration of parent drug and hepatic/enteric-derived metabolites in the urine as a result of renal excretion. Breaching of the barrier provided by the bladder epithelial lining (the urothelium) can expose the underlying tissues to urin...

In vivo studies of implanted acellular biological scaffolds in experimental animals have shown constructive remodelling mediated by anti-inflammatory macrophages. Little is known about the human macrophage response to such biomaterials, or the nature of the signalling mechanisms that govern the macrophage phenotype in this environment. The cellular...

This chapter reviews recent developments for novel regenerative medicine approaches for urinary bladder reconstruction. The chapter introduces clinical requirements for functional tissue replacement and discusses the use of synthetic and natural matrices for bladder reconstruction. It then describes the application of cell-seeded bio-matrices using...

Despite major advances in high-throughput and computational modelling techniques, understanding of the mechanisms regulating tissue specification and differentiation in higher eukaryotes, particularly man, remains limited. Microarray technology has been explored exhaustively in recent years and several standard approaches have been established to a...

Tissue-engineering and regenerative medicine strategies for the bladder and urinary tract are dependent on the ability to generate adequate numbers of differentiation-competent uro-epithelial cells. In situ, urothelium is a mitotically-quiescent, but highly regenerative epithelium. Although evidence supports a resident, basally-located urothelial p...

There is an emerging association between ketamine abuse and the development of urological symptoms including dysuria, frequency and urgency, which have a neurological component. In addition, extreme cases are associated with severe unresolving bladder pain in conjunction with a thickened, contracted bladder and an ulcerated/absent urothelium. Here...

The transcription factor octamer-binding protein 4 (Oct4; encoded by POU5F1) has a key role in maintaining embryonic stem cell pluripotency during early embryonic development and it is required for generation of induced pluripotent stem cells. Controversy exists concerning Oct4 expression in somatic tissues, with reports that Oct4 is expressed in n...

Calcium signalling plays a central role in regulating a wide variety of cell processes. A number of calcium signalling models exist in the literature that are capable of reproducing a variety of experimentally observed calcium transients. These models have been used to examine in more detail the mechanisms underlying calcium transients, but very ra...

Viable human bladder tissue is an essential resource for cell biological research aimed at understanding human diseases of the bladder and for developing new Tissue Engineering and Regenerative Medicine (TERM) strategies for bladder reconstruction. Typically, normal human urinary tract tissue is obtained from adult or paediatric surgical patients w...

Further to its role as a physical barrier, the urothelium is considered to play an active role in mechanosensation. A key mechanism is the release of transient mediators that activate purinergic P2 receptors and transient receptor potential (TRP) channels to effect changes in intracellular calcium. Despite the implied importance of these receptors...

Scratch wounding of a urothelial cell monolayer triggers a number of events including the release of soluble, diffusible signalling factors and mechanical stimulation of cells at the wound edge. These events cause a sustained elevation in cytosolic calcium concentration in the cells surrounding the wound and a transient rise in those further away....

Transforming growth factor (TGF) β has diverse and sometimes paradoxical effects on cell proliferation and differentiation, presumably reflecting a fundamental but incompletely-understood role in regulating tissue homeostasis. It is generally considered that downstream activity is modulated at the ligand:receptor axis, but microarray analysis of pr...

Influence of medium supplements on the basal phosphorylation of SMAD3 in NHU cell cultures. NHU cells were grown in KSFMc or KSFM with no supplements, with BPE alone (60 µg/ml) or EGF alone (6 µg/ml), all in the presence or absence of SB431542 (3 µM). KSFMc with TGF-β1 (2 ng/ml) was used as a positive control for pSMAD3 activation. Cell lysates (25...

Expression of TGFβ ligands and probes taken from analysis of gene chip data. P = present; A = absent (DOC)

Verification that differentiated NHU cell cultures used for Affymetrix arrays expressed established urothelial differentiation-associated genes. Analysis of marker gene expression from arrays performed at 144 h post induction of differentiation by ABS/Ca2+ and TZ/PD protocols compared to the autologous 24 h non-differentiated control culture. Resul...

To develop a robust sterile, fully demucosalized and vascularized seromuscular patch for use as an adjunct to novel bioengineering techniques aimed at augmenting, reconstructing, or replacing the bladder because of endstage disease. To eliminate deep colonic epithelial crypts to prevent the possibility of colonocyte regrowth. To maintain sterility...

Malignant development cannot be attributed alone to genetic changes in a single cell, but occurs as a result of the complex interplay between the failure of cellular regulation mechanisms and the presence of a permissive microenvironment. Although E-cadherin is classified as a 'metastasis suppressor' owing to its role in intercellular adhesion, the...

It is generally considered that the bladder is impervious and stores urine in unmodified form on account of the barrier imposed by the highly-specialised uro-epithelial lining. However, recent evidence, including demonstration of aquaporin (AQP) expression by human urothelium, suggests that urothelium may be able to modify urine content. Here we ha...

Growing evidence indicates that Rab GTPases, key regulators of intracellular transport in eukaryotic cells, play an important role in cancer. We analysed the deregulation at the transcriptional level of the genes encoding Rab proteins and Rab-interacting proteins in bladder cancer pathogenesis, distinguishing between the two main progression pathwa...

Genes deregulated during bladder cancer pathogenesis: second set of data. Second set of data. Affymetrix HG U95A/U95Av2 DNA microarrays. Genes down- or up-regulated in the tumor samples. Left: FGFR3-non-mutated tumor groups. Right: FGFR3-mutated tumor groups. The results shown to pass the thresholds: FC>1.5 (or <0.667) and qValue <5% with the first...

Pearson correlation (r) (and pValue) between the expression of MKI67 and the expression of genes listed in left column. Pearson correlation (r) (and pValue) between the expression of MKI67 and the expression of genes listed in left column in 28 Ta G1G2 (FGFR3-mutated) tumor samples and 63 T2-4 (FGFR3-non-mutated) tumor samples. Are highlighted (in...

Pearson correlation (r) (and pValue) between the expression of different urothelial differentiation markers genes and the expression of genes listed in left column. Pearson correlation (r) (and pValue) between the expression of FOXA1, GRHL3, UPK1A/B, UPK2, UPK3A/B and the expression of genes listed in left column in 28 Ta G1/G2 (FGFR3-mutated) tumo...

Rab clusters. A Rab cluster is defined as a Rab and its interacting proteins: the GEFs (guanine nucleotide exchange factors), the GAPs (GTPase activating proteins) and the effector proteins. Rab proteins are in yellow, GEFs in green, GAPs in light red and effector proteins in blue. (PDF)

List of papers in which the Rab partners were originally described. (XLS)

Genes significantly up- or down-regulated in each Rab cluster for each tumor group of the two pathways. “D” (green) corresponds to a gene significantly down-regulated in the tumor samples, “U” (red) corresponds to a gene significantly up-regulated in the tumor samples. “% All genes” corresponds to the total number of genes up- or down-regulated wit...

Genes deregulated only in FGFR-non-mutated (A) and mutated tumors (B). Genes up- (or down)-regulated that pass the thresholds: FC>1.5 (or <0.667) and qValue <5% only for the FGFR3-non-mutated (A) or FGFR3-mutated (B) tumor pathway. Left column: comparison with the normal urothelium samples. Right column: comparison with the tumor samples of same st...