Heterogeneous components of lung adenocarcinomas harbour distinct EGFR mutations and PD-L1 expression
A recent study using laser capture microdissection demonstrated that different areas of the same lung adenocarcinoma tissue can harbour both sensitising EGFR mutations and those mutations that confer resistance to TKIs.
Reducing intestinal inflammation with secondary bile acids
Intestinal inflammation was shown to be associated with reduced levels of secondary bile acids and bacteria of the Ruminococcaceae family in a recent study moving onto clinical trials.
Probiotic Bacillus subtilis protects against α-synuclein aggregation in C.elegans
A new study looking at B.subtilis in C.elegans could open the possibilities of diet-based intervention and the development of protective bacterial metabolites to combat Parkinson's disease.
Flagellin-elicited adaptive immunity suppresses flagellated microbiota and vaccinates against chronic inflammatory diseases
A recent study has demonstrated positive in vivo results for a flagellin-based vaccine for IBD treatment.
ATR kinase inhibitor AZD6738 potentiates CD8+ T cell-dependent antitumour activity following irradiation
Authors of a recent study have shown that the ATR kinase inhibitor combined with local tumour radiation generated a durable CD8+ T cell-dependent anti-tumour response.
Regulatory network analysis of Paneth cell and goblet cell enriched gut organoids using transcriptomics approaches
A recent study used cell-enriched organoids in order to predict the key molecular regulators involved in Paneth cell and goblet cell specific functions.
Epithelial CD47 is critical for mucosal repair in the murine intestine in vivo
Researchers have demonstrated an essential role of CD47 in promoting cell migration in a recent wound healing study.
Fluorescence of Picrosirius Red Multiplexed with Immunohistochemistry for the Quantitative Assessment of Collagen in Tissue Sections
An emerging method of imaging offers a new opportunity to quantify collagen around areas of interest and an innovative approach to study the role of collagen in controlling cell behaviour.
The use of organoids to study the epithelial response to secreted pro-inflammatory cytokines
A recent organoid study suggests that Nfkb2 signalling pathway may be targetable to improve intestinal inflammation.
Genome-wide epigenetic and transcriptomic characterisation of human-induced pluripotent stem cell-derived intestinal epithelial organoids
Recent developments have allowed us to generate epithelial cell-specific intestinal organoids from pluripotent stem cells, we review a study which aims to characterise these organoid cultures and compare them with cultures derived from primary intestinal epithelial cells.
5-Fluorouracil treatment induces characteristic T>G mutations in human cancer
Organoid models treated with 5-FU suggest further oncology scrutiny is needed when treating younger patients
A murine platform to evaluate therapies against radiation-induced gastrointestinal syndrome
Models to service radiation-induced gastrointestinal syndrome are limited, this study shows a platform to study genes linked to intestinal radiation injury
Plucked hair follicles from patients with Chronic Discoid Lupus Erythematosus (CDLE) display a disease-specific genomic molecular signature
This pilot study showed that a disease-specific gene signature for Chronic Discoid Lupus Erythematosus can be obtained from plucked hair follicles followed by gene expression analysis.
In vivo selection reveals autophagy promotes adaptation of metastatic ovarian cancer cells to abdominal microenvironment
An increase in autophagic potency among ovarian cancer cells may be crucial for selection of metastatic colonies in the abdominal microenvironment. Autophagy may be a promising potential therapeutic target in the treatment of ovarian cancer.
Derivation of adult canine intestinal organoids for translational research in gastroenterology
This study describes the development and long-term maintenance of small intestine and colon in vitro canine organoids using whole tissue and biopsy samples from 40 healthy and diseased cohorts.
Spatial metagenomic characterisation of microbial biogeography in the gut
Evaluation of a novel technique, MaPS-seq, a multiplexed protocol combining the positive aspects of metagenomics, transcriptomics and imaging to analyse the gut microbiome.
Formaldehyde scavenging agents and the chemistry of antigen retrieval in FFPE tissues, and their potential for improved development of IHC staining protocols
A new approach to identify new and effective Antigen Retrieval reagents based on their ability to scavenge previously dissociated formaldehyde.
Measuring intratumoral vessel density using automated image analysis
Digital image analysis to analyse the intratumoral vessel density within specified regions of resected colon and TMA sections of colon tumour.
The importance of Reg4 in the response of the colonic epithelium to inflammation
Upregulation of, or treatment with Reg4 was associated with the maintenance and proliferation of colonic organoid cultures, the epithelial response to mucosal injury in an in vivo model of chemically-induced acute colitis (DSS) and in paediatric patients with Intestinal Failure and the response of intestinal epithelial cells to an inflammatory stimulus (LPS) in vitro.
Human 3D Gastrointestinal Microtissue Barrier Function As a Predictor of Drug-Induced Diarrhoea
New preclinical approaches are required for earlier testing of therapies for GI-toxicity. In this study the authors describe an in vitro human 3D GI microtissue and validated it for GI-toxicity using a panel of known drugs that do/do not cause diarrhoea.
Identification of a Radiosensitivity Molecular Signature Induced by Enzalutamide in Hormone-sensitive and Hormone-resistant Prostate Cancer Cells
Androgen Deprivation Therapy (ADT) has demonstrated improved clinical success with further anti-tumour efficacy when combined with XRT, however, half of patients develop a resistance. In this study the authors identified potential prognostic biomarkers for response to combined ENZA, ADT and XRT therapy.
Prophylactic TNF blockade uncouples efficacy and toxicity in dual CTLA-4 and PD-1 immunotherapy
Prophylactic TNF blockade may safely allow doses of ipilimumab to be increased in immune checkpoint blockade combined therapies.
Defining disease mechanism and probing for effective treatments for Generalized Pustular Psoriasis by RNA-seq transcriptomics profiling
To investigate GPP pathogenesis and the efficacy of drugs on molecular level, Wang et al performed RNA-sequencing on peripheral blood mononuclear cells (PBMCs) obtained from five adult GPP patients before and during treatment with the oral retinoid acitretin.
The delayed effects of acute radiation exposure within the mouse intestine
Partial-body irradiation (PBI) models with minimal (2.5% or 5%) Bone Marrow (BM) sparing results in Acute GI Damage (GI-ARS). However, it enables longer-term survival against radiation levels that would normally be lethal due to BM Toxicity enabling us to study the long-term effects of exposure.
Distinct Immune Cell Populations Define Response to Anti-PD-1 Monotherapy and Anti-PD-1/Anti-CTLA-4 Combined Therapy
To discover potential biomarkers of response and resistance to anti-PD1 monotherapy and combined anti-PD1/anti-CTLA-4 immunotherapies in a cohort of melanoma patients.
Triggering of a Dll4
Diabetic foot ulceration (DFU), a chronic diabetes complication, represents a major medical, social and economic problem. This study identified a Notch1 signaling pathway as a crucial mediator of impaired wound healing in diabetes.
Next Generation Sequencing versus Microarray
Epistem are pleased to announce their acquisition of the Illumina NextSeq 550 next generation sequencer. With tunable output and high data quality, it provides the flexible power you need for whole-genome, transcriptome, and targeted resequencing.
Hedgehog stimulates hair follicle neogenesis by creating inductive dermis during murine skin wound healing
Mammalian adult skin wounds typically heal by fibrotic repair without hair follicle (HF) regeneration. This study is the first to show that de novo HFs can be regenerated in adult skin by modulating a Shh pathway in the dermis.
Redefining the practical utility of blood transcriptome biomarkers in inflammatory bowel disease
Screening for IDB candidate genes using RNA sequencing of whole blood liquid biopsies taken from relatively large cohorts of both adult and pediatric patients with defined clinically active IBD.
Epithelial-mesenchymal transition in human prostate cancer demonstrates enhanced immune evasion marked by IDO1 expression
Cancer invasion and metastasis are driven by epithelial-to-mesenchymal transition (EMT). This study identifies an important link between EMT and immune evasion in prostate cancer patients.
Overexpression of miR-221 and miR-222 in the cancer stroma is associated with malignant potential in colorectal cancer
In this study the authors used laser capture microdissection to investigate the role of stromal miRNA in patients with metastatic cancer, aiming to identify those miRNAs present in the stroma that may contribute to metastases.
Transcriptomic analysis of hepatocellular carcinoma reveals molecular features of disease progression and tumour immune biology
Hepatocellular carcinoma (HCC) is a common cause of cancer deaths with a particularly poor prognosis. A recent publication by Okrah et al., describing molecular aspects of HCC progression illustrates the power of multigene transcriptomic signatures suggesting a rationale for immunotherapy early in HCC disease progression and potentially in combination with an anti-fibrotic strategy.
The role of immunohistochemistry in patient stratification and personalised medicine
Not all patients respond well to treatment and drugs have failed in the clinic due to “all patient” designs. Immunohistochemistry (IHC) could be used to group patients according to the expression levels of a specific biomarker. With successful validation and quantification, IHC assays can play a vital role in drug development as a method of patient stratification.
A stromal cell population that inhibits adipogenesis in mammalian fat depots
Adipocyte development and differentiation play an important role in the origin of obesity and many related diseases such as type 2 diabetes and cancer. Using a single-cell transcriptomics-based approach in a mouse model revealed three different adipocyte progenitor cell populations, including one subpopulation, termed Aregs, that suppressed adipocyte formation in vitro and in vivo.
Acute Lymphoblastic Leukaemia highjacks a neural mechanism to invade the CNS
Acute Lymphoblastic Leukaemia (ALL) most often metastasise to organs such as the spleen and lymph nodes, but also to the Central Nervous System (CNS). Here, Yao et al show that ALL cells infiltrate the CNS not by breaching the blood-brain barrier, but by hijacking the neural migratory pathways.
Determinants of variability of five programmed death ligand-1 immunohistochemistry assays in non-small cell lung cancer samples
Upregulation of PD-L1 is associated with increased risk and tumourigenicity of cancers, helping many escape immune surveillance. PD-1/PD-L1 expression has been suggested as a possible biomarker for identifying patients that may be sensitive to PD1/PD-L1 immunotherapies. Current PD1/PD-L1 testing procedures may not be sufficient to distinguish responders and this study aimed to simplify the process by assessing the different methods on clinically-relevant NSCLC samples, to identify factors relevant for PD-L1 detection and make assays more robust and reproducible.
Gene expression profiles from plucked scalp hair to assess pharmacodynamic biomarkers following treatment of advanced solid tumors with a novel anti-cancer stem cell Wnt inhibitor
Wnt signaling pathway genes are crucial for cell fate determination and cell polarity during development. Ipafricept (OMP-54F28) is a first in class recombinant fusion protein producing a truncated decoy receptor that binds to a cystine-rich region of Wnt ligand and as a result it blocks the activation of the Wnt signaling. Here the authors described the results of a phase 1 first in-human clinical trial.
Improved healing of large wounds via the reprogramming of wound-resident mesenchymal cells into epithelial cells
The authors of this paper demonstrated that mesenchymal cells within a large wound could be converted into keratinocytes in vivo via the transduction of four genes. This improved re-epithelialization of wounds through generation of new epithelial tissue that showed normal morphology/characteristics and eventually connected to host skin. The paper provides proof of principle for a new wound healing treatment particularly useful for large wounds that normally require skin grafts.
The influence of TLR5 expression by intestinal epithelial cells on the gut microbiome
It has been shown that TLR5 expression by the intestinal epithelium early in neonatal development is an important factor in preventing increased colonisation by flagellated bacteria, which can drive chronic pathologies such as metabolic syndrome and colitis
Two recent papers highlight the importance of tumour microenvironment on cancer cell stemness
Two recent papers demonstrate the importance of the tumour microenvironment on its progression through in vitro co-culture. The authors in one paper show that M2 macrophages increase the stem cell capacity of an ovarian cancer cell line, whilst in the other fibroblasts increase the stemness of a lung carcinoma cell line. They highlight the importance of considering the tumour microenvironment in in vitro experimentation, and also reveal potential new therapeutic targets.
CAR-T Cells in cancer Therapy
Summary of a paper reporting an unusually delayed response from one patient during a clinical trial in CLL using CAR T-cells targeted to CD-19 that may shed light on some of the mechanisms determining persistence of CAR T-cells and their clinical outcome.
Memory of EpSC in Wound Healing
Summary of a study investigating how skin inflammation effects EpSCs to allow them to speed up barrier restoration following subsequent tissue damage.
4PD1hi cells predict response to immunotherapy
An interesting paper published in Cancer Cell explores the role of a newly identified and non-conventional subset of inhibitory CD4+ T cells.
Can changes to gut microbiota affect the efficacy of chemotherapies?
Can changes to gut microbiota affect the efficacy of chemotherapies?
Oncogenic JAK2V617F causes PD?L1 expression, mediating immune escape in myeloproliferative neoplasm
Oncogenic JAK2V617F causes PD-L1 expression, mediating immune escape in myeloproliferative neoplasms
Glut1 - A Target for Psoriasis Treatment
Targeting breast cancer with Hsp70?aptamer?guided nanoparticles with chemotherapeutic payloads
Targeting breast cancer with Hsp70-Aptamer-guided nanoparticles with chemotherapeutic payloads
The relationship between epithelial cell fate, microbiota composition and inflammation in the intestine
How perturbation of the homeostatic mechanisms governing epithelial cell fate in the intestine influence the gut microbiota and the susceptibility to colitis. The influence of structural proteins on key signalling pathways and their downstream transcriptional machinery are discussed.
CAR-T Cells: Engineering Immune Cells to Treat Cancers
For many years surgery, chemotherapy and radiation therapy have been the foundations of cancer treatment. Over the last two decades, targeted therapies have cemented themselves as standard treatments for many cancers and over the past several years, immunotherapy therapies that enlist and strengthen the power of a patient’s immune system to attack tumours has emerged.
Epistem Analytical Protein Services
Epistem offer protein analysis as a vital readout of Test Item efficacy in vivo and in vitro. At Epistem, we offer 2 automated platforms that can be used to analyse proteins in cells and liquid samples - flow cytometry and Luminex
Laser Capture Microdissection (LCM) - A Strategy for Success
Micro-sampling by laser capture microdissection is a powerful tool allowing specific analysis from whole tissues, in heterogeneous microenvironments such as tumours, where less well represented target cells are dwarfed by tissue stroma and other cell types. Whilst the application is potentially powerful for target discovery and mechanistic understanding, the process is notoriously difficult, particularly in clinical specimens. Our Laser Capture Microdissection team has evaluated the findings from all the studies run in our GCLP-accredited labs and devised a strategy for LCM studies maximising the potential of success and giving the sponsor control over go/no go decisions.
New orthotopic organoid xenograft model enables reconstruction of human colon in vivo
A recent Cell Stem Cell paper describes a novel technique to orthotopically engraft human colon organoids in vivo. They observed long-term full reconstruction of the human colon upon engraftment, self-renewal of human Lgr5+ stem cells and multi-lineage differentiation. This model allows investigation of human stem cell functionality. Indeed, data presented demonstrates cell-intrinsic control of proliferation and differentiation within the human tissue, with slower rates of stem cell cycling observed than that of the host.
Stratification of Patients using Gene Expression Analysis of Laser Capture Microdissected (LCM) Tissue and Histology/IHC
This procedure could help stratify patients where pre-treatment could be avoided, these patients may then benefit from advanced surgery or alternative treatment options and prevent unnecessary toxicities.
CCDC88B and its Importance in Inflammatory Bowel Disease
Epistem have extensive experience of running in vivo colitis models including off-the-shelf models of adoptive T cell transfer-mediated and DSS-induced acute colitis; in addition, we are happy to develop and run bespoke models in collaboration with Sponsors.
Choosing the best preclincal oral mucositis model to demonstrate efficacy or Mechanism of Action
The oral mucosa is a stratified squamous epithelium that has a crucial barrier function. Any damage or loss of integrity during oncology therapies poses a major risk of infection, since such patients are often also immunocompromized. Furthermore, the large ulcerations can be so painful that patients are often unable to eat and require hospitalization and enteral nutrition.
Antibody Validation: Does the antibody really recognise what it says on the label?
Epistem has an expert histology lab whose capabilities are available as a standalone service, or integrated to add value to your preclinical model. Nicola Tonge, Epistem's Histology Manager and IHC specialist discusses the issues faced when choosing the correct antibody as there is currently no industry standard for antibody validation, and not all antibodies work 'how they say on the label'.
Epistem Bioinformatics Services
Epistem's pharmacogenomics division have provided biomarker discovery and validation studies with many sponsors. The bespoke nature of many of Epistem's services mean that bioinformatics analysis is always tailored to the nature of the study.
Gene expression analyses of murine skin highlight strain-dependent effects of Imiquimod on psoriatic signatures
Our gene expression analysis and detailed protein analyses by flow cytometry and Luminex
JAK2, a novel therapeutic target in lung adenocarcinoma?
The field of personalised medicine to provide understanding of the molecular basis of disease is constantly growing. Multiple signalling pathways have been identified that associate with malignant transformation. However, the vast majority of lung cancer patients have no known driver genes detected, and they are still treated with standard cytotoxic chemotherapy.
Can organoids guide clinical decisions? Current successes and potential pitfalls
A paper in the Nature Medicine journal reviews the use of organoids in the clinic up-to-date and also highlights some of the potential obstacles for widespread use as predictive tools for in vivo treatment. Our Senior Scientist, Dr Sarah Hoyle, summarises the key points from this paper.
Platelets are major suppressors of T cell function
We currently offer validated models to address immune function in tumour bearing mice. Our detailed flow cytometric expertise can further assess the identity of tumour infiltrating and peripheral cell populations such as CD8+ T cells, MDCSs and regulatory T cells, by surface identification of key molecules, or the intracellular mediators they express (cytokines, transcription factors).
Human organoids provide new insights into early stage enterovirus infection
Epistem have presented developmental work within the organoid model in the areas of epithelial regeneration and GI toxicity at various conferences. Potential readouts in this model could be proliferation, differentiation, apoptosis, cytotoxicity screening and protein and gene expression.
Adoptive T Cell Transfer Models of Colitis
This model offers the ability to study test items using a range of dosing regimens, from dosing in a prophylactic manner through to examining efficacy in ameliorating early stage or established disease.
CCL20 as a biomarker of drug efficacy in IBD
The chemokine CCL20 (Mip-3?) plays an important role in recruiting Th17 cells (which express the corresponding receptor, CCR6) to sites of inflammation, thus playing a role in the pathogenesis of chronic inflammatory conditions such as Crohn's disease. In a recent paper in the Journal of Crohn's and Colitis, Giovanni Monteleone and colleagues presented data from clinical and laboratory studies on Mongersen, a Smad7 antisense oligonucleotide.
Development of a Signature of Aberrant Myc Transcription has Prognostic Value in Evaluating Novel Myc-Targeted Therapies
Altered expression of transcription factors are key indicators in cancer progression in functions such as proliferation and differentiation, angiogenesis and apoptosis. A significant proportion of cancers are driven by mutations in these networks, including Wnt/?-catenin, Ras/MAPK and Myc
A new paper demonstrates that BMP limits Lgr5+ stem cell number in intestinal epithelial cells
The paper demonstrates that BMP acts via Smad to transcriptionally repress a range of stem cell associated genes, including Lgr5, and that this is independent of the Wnt//??catenin signalling pathway.
A new opportunity to reduce scarring by converting myofibroblasts to adipose cells?
Epistem has validated in vivo excisional and incisional, human skin engraft, diabetic and LPS-induced inflammation wound healing models. In vitro wound healing assays are also available to assess cell proliferation/migration, wound closure and angiogenesis.
On-going Mechanical Damage from Mastication Drives Homeostatic Th17 Cell Responses at the Oral Barrier
We currently offer in vivo models of rheumatoid arthritis, psoriasis and inflammatory bowel disease.
Accessing Disease End-Points in IBD Models
High quality histopathology analysis is standard for all our IBD service models. In addition, Epistem offer a wide range of analytical services that allow Sponsors to acquire quantitative experimental data for a variety of disease-relevant end-points.
Laser Capture Microdissection for Gene Expression Analysis in Distinct Cell Populations
Knowledge in early phase cancer development to provide understanding of the mechanism of action of oncogenic mutations is constantly growing. A recent study carried out by Mo et al has highlighted several interesting results.
Organoids programmed to differentiate into rare hormone-producing enteroendocrine cells
New paper describes protocols for the manipulation of organoid Lgr5+ stem cells into quiescence before being programmed to differentiate into specific intestinal lineages, including the rare hormone-producing endocrine cell
New paper demonstrating the microbiota impact on cell proliferation via microRNA expression changes in intestinal epithelial stem cells
Intestinal flora are known to impact the immune system and metabolism, with chronic exposures to certain flora linked to conditions such as inflammatory bowel disease and fibrosis. Breeches of the epithelial barrier and systemic infection can lead to more acute problems such as sepsis.
New biomarker/therapeutic target in AML
The field of personalised medicine to provide understanding of the molecular basis of disease and the mechanism of action of chemotherapeutic drugs is constantly growing. A recent study carried out by Schneider et al has highlighted several interesting facts.
Citrulline as a Biomarker of GI Toxicity
Citrulline is produced by small bowel enterocytes, with circulating citrulline levels correlating with enterocyte mass. Changes in enterocyte number (or in some cases function) are therefore reflected in plasma levels. As a result, citrulline levels may be a biomarker for various GI syndromes
The Tumor Microenvironment Represses T Cell Mitochondrial Biogenesis to Drive Intratumoral T Cell Metabolic Insufficiency and Dysfunction
We currently offer 3 models to address the immune response in tumour bearing mice. Metabolic and mitochondrial function of T cells can be assessed by flow cytometry. Our detailed flow cytometric expertise can further assess the identity of tumour infiltrating and peripheral cell populations, by surface identification of key molecules, or the intracellular mediators they express (cytokines, transcription factors).
Multiplexing Immunohistochemistry: A new method for integrating biomarker expression
Multiplexed biomarkers and diagnostics can provide a greater understanding of the response of a tissue to a drug, or of disease stage, allowing further progress towards personalised medicines.
Novel drug prevents AML cells from hiding
Plucked hair ex vivo model identifies novel Notch PD biomarker suitable for use in the clinic.
Epistem have validated plucked scalp hair as a robust ex vivo model allowing biomarker signature identification. Readouts in these models include expression profiling by microarray or RNA-Seq, and targeted assays by qPCR.
FDA approval for new diabetic wound dressing.
Epistem validated in vivo diabetic wound healing model showing delayed wound healing (wound closure and re-epithelisation) and a decreased inflammatory response.
New insights into epithelial barrier dysfunction in the intestine: relevance for IBD
Alterations in the complex inter-relationship between intestinal epithelial cells, the mucosal immune system responses and the gut microbiota are responsible for the development of inflammatory bowel disease in humans.
Novel sunscreen ingredient demonstrates high protection from UVA radiation
Epistem have validated 3D and 2D skin models. 3D models include organ cultures and the human living skin equivalent that closely resembles fully stratified human epidermis with a dermal region. 2D models include monolayer cultures of primary epithelial and dermal cell types.