Hippokratis Kiaris

Improvement of chemotherapeutic drug efficacy by endoplasmic reticulum stress

Tunicamycin (TUN), an inhibitor of protein glycosylation and therefore a potent stimulator of endoplasmic reticulum (ER) stress, has been used to improve anticancer drug efficacy, but the underlying mechanism remains obscure. In this study, we show that acute administration of TUN in mice induces the unfolded protein response and suppresses the levels of P21, a cell cycle regulator with anti-apoptotic activity. The inhibition of P21 after ER stress appears to be C/EBP homologous protein (CHOP)-dependent because in CHOP-deficient mice, TUN not only failed to suppress, but rather induced the expression of P21. Results of promoter-activity reporter assays using human cancer cells and mouse fibroblasts indicated that the regulation of P21 by CHOP operates at the level of transcription and involves direct binding of CHOP transcription factor to the P21 promoter. The results of cell viability and clonogenic assays indicate that ER-stress-related suppression of P21 expression potentiates caspase activation and sensitizes cells to doxorubicin treatment, while administration of TUN to mice increases the therapeutic efficacy of anticancer therapy for HepG2 liver and A549 lung cancers.

Regulation of P21 during diabetes-associated stress of the endoplasmic reticulum

Endoplasmic reticulum (ER) stress plays a major role in the pathogenesis of diabetes by inducing β-cell apoptosis in the islets of Langerhans. In this study, we show that the transcription factor CHOP, which is instrumental for the induction of ER-stress-associated apoptosis and the pancreatic dysfunction in diabetes, regulates the expression of P21 (WAF1), a cell cycle regulator with anti-apoptotic activity that promotes cell survival. Deficiency of P21 sensitizes pancreatic β-cells to glucotoxicity, while in mice genetic ablation of P21 accelerates experimental diet-induced diabetes, results indicative of a protective role for P21 in the development of the disease. Conversely, pharmacological stimulation of P21 expression by nutlin-3a, an inhibitor of P53-MDM2 interaction, restores pancreatic function and facilitates glucose homeostasis. These findings indicate that P21 acts as an inhibitor of ER-stress-associated tissue damage and that stimulation of P21 activity can be beneficial for the management of diabetes and probably of other conditions in which ER-stress-associated death is undesirable.

Inhibition of CDK8 mediator kinase suppresses estrogen dependent transcription and the growth of estrogen receptor positive breast cancer

Hormone therapy targeting estrogen receptor (ER) is the principal treatment for ER-positive breast cancers. However, many cancers develop resistance to hormone therapy while retaining ER expression. Identifying new druggable mediators of ER function can help to increase the efficacy of ER-targeting drugs. Cyclin-dependent kinase 8 (CDK8) is a Mediator complex-associated transcriptional regulator with oncogenic activities. Expression of CDK8, its paralog CDK19 and their binding partner Cyclin C are negative prognostic markers in breast cancer. Meta-analysis of transcriptome databases revealed an inverse correlation between CDK8 and ERα expression, suggesting that CDK8 could be functionally associated with ER. We have found that CDK8 inhibition by CDK8/19-selective small-molecule kinase inhibitors, by shRNA knockdown or by CRISPR/CAS9 knockout suppresses estrogen-induced transcription in ER-positive breast cancer cells; this effect was exerted downstream of ER. Estrogen addition stimulated the binding of CDK8 to the ER-responsive GREB1 gene promoter and CDK8/19 inhibition reduced estrogen-stimulated association of an elongation-competent phosphorylated form of RNA Polymerase II with GREB1. CDK8/19 inhibitors abrogated the mitogenic effect of estrogen on ER-positive cells and potentiated the growth-inhibitory effects of ER antagonist fulvestrant. Treatment of estrogen-deprived ER-positive breast cancer cells with CDK8/19 inhibitors strongly impeded the development of estrogen independence. In vivo treatment with a CDK8/19 inhibitor Senexin B suppressed tumor growth and augmented the effects of fulvestrant in ER-positive breast cancer xenografts. These results identify CDK8 as a novel downstream mediator of ER and suggest the utility of CDK8 inhibitors for ER-positive breast cancer therapy.

A CCL8 gradient drives breast cancer cell dissemination.

The migration of cancer cells towards gradients of chemoattractive factors represents a potential, yet elusive, mechanism that may contribute to cancer cell dissemination. Here we provide evidence for the maintenance of a gradient of increasing CCL8 concentration between the epithelium, the stroma and the periphery that is instrumental for breast cancer cells’ dissemination. In response to signals elicited by the neoplastic epithelium, CCL8 production is enhanced in stromal fibroblasts at the tumor margins and in tissues at which breast cancer cells tend to metastasize such as the lungs and the brain. Manipulation of CCL8 activity influences the histology of the tumors and promotes major steps of the metastatic process such as invasion to adjacent stroma, intravasation and ultimately extravasation and seeding. These findings exemplify how gradients of chemoattractive factors such as CCL8, drive metastasis and suggest that interference with their operation may provide means for breast cancer management.

Crosstalk between C/EBP homologous protein (CHOP) and glucocorticoid receptor in lung cancer.

Loss of homeostasis triggers the endoplasmic reticulum (ER) stress response and activates the unfolded protein response (UPR) resulting in the induction of the CCAAT/enhancer binding protein (C/EBP) homologous protein (CHOP). Glucocorticoids (GCs), via the glucocorticoid receptor (GR), regulate numerous physiological processes in an effort to maintain homeostasis. Previous studies demonstrated that glucocorticoids suppress ER stress by enhancing correct folding of secreted proteins and degradation of misfolded proteins. Here, we describe a novel crosstalk between ER-stress and the glucocorticoid receptor signaling. We showed that treatment of wild type mice with Tunicamycin (inducer of ER-stress) increased GR protein levels in the lungs. Treatment of A549xa0cells (human lung cancer cells) with ER stress inducers modulated the Dexamethasone-induced subcellular localization of GR and the phosphorylated forms of GR (pGRSer211 and pGRSer203) with concomitant changes in the expression of primary GR-target genes. We demonstrated a significant protein-protein interaction between GR and CHOP, (GR-CHOP heterocomplex formation) under ER stress conditions. The functional consequences of ER stress- GR signaling crosstalk were assessed and demonstrated that long time exposure (24-48xa0h) of A549xa0cells to dexamethasone (10(-6)xa0M) reversed the Tunicamycin-induced cell death, a phenomenon associated with parallel increases in GR protein content, increases in cell survival parameters and decreases in cell apoptosis-related parameters. Our study provides evidence that there is a cross talk between ER-stress and GR signaling, this being associated with mutual functional antagonism between CHOP and GR-mediated pathways in lung cells with important implications in lung cell function.

Modeling estrogen receptor-positive breast cancers in mice: is it the best we can do?

The continuous supplementation of mice with supraphysiological doses of estrogen u2028for the growth of estrogen receptor-positive breast cancers has been linked to toxicity in the host and perturbation of cancer cells function that can misguide preclinical studies. Thus, alternative experimental models with circulating levels of estrogens higher than those of mice may represent more suitable hosts to model estrogen receptor-positive breast cancers.

The challenges faced by living stock collections in the USA

Many discoveries in the life sciences have been made using material from living stock collections. These collections provide a uniform and stable supply of living organisms and related materials that enhance the reproducibility of research and minimize the need for repetitive calibration. While collections differ in many ways, they all require expertise in maintaining living organisms and good logistical systems for keeping track of stocks and fulfilling requests for specimens. Here, we review some of the contributions made by living stock collections to research across all branches of the tree of life, and outline the challenges they face. DOI: http://dx.doi.org/10.7554/eLife.24611.001

Endoplasmic Reticulum stress is associated with the pathogenesis of Pemphigus Vulgaris

tion of Met during bleaching (s3). Trp, an amino acid that is susceptible to reactive oxygen species (s4) or thermal reaction (5), was depleted by the chemical treatments of hair. In addition to the individual alteration of these AAs, multivariate analysis using PCA revealed that combined assessment of Trp, Met, Cya and Cys/Cya may be useful in grading hair damage associated with chemical treatments. High levels of Trp, Met and Cys/ Cya suggested intact or healthy hairs, while their decreases indicated hair damage. Along with the recently reported proteomic approach (6), this methodology may overcome the limitation of morphological or mechanical evaluation by providing information on the compositional changes, which is crucial to the prevention or repairing of damaged hairs. Hair lipids are important components of hair cuticle. Hair lipids are mainly composed of FFAs, CH, cholesteryl sulphate, wax esters, 18-MEA, triglyceride and CERs (7). Of these hair lipids, FFAs like C22:1FA (erucic acid), C22FA (behenic acid), C24FA (lignoceric acid), C24:1FA (nervonic acid), C26FA (cerotic acid) and an integral hair lipid, 18-MEA, were depleted in proportion to the severity of hair damage. FFAs originated from sebum secretion coat the outside of hairs (8). Therefore, it is plausible that these FFAs are easily depleted during chemical treatment of hair. 18-MEA, which is biosynthesized in hair matrix cells, is linked by a cysteine thioester bond within the cuticle at the surface of human hairs (s5), which makes it susceptible to the breakage of cross-links during chemical treatment as well as in diseases (9). In conclusion, we demonstrate that hair AAs and lipids are markedly altered during chemical treatments of hair. The alteration of these AAs and lipids that are critical to impart mechanical strength and barrier function on hair may ultimately deteriorate the integrity or fibre quality of hair. Metabolomic analysis revealed AA or lipid biomarkers specific to hair damage, which may be useful for the quantitative grading of hair damage, and ultimately would be instrumental for the development of safer and undamaging hair care products. Author contributions KMJ, BMK, HKL, JHL and KML designed the study; KMJ, ARK and SNK involved in the research; KMJ, KML and SJB analysed the data; and KMJ and KML wrote and revised the manuscript. Conflict of interest None Supporting Information

Growth of human breast cancers in Peromyscus

ABSTRACT Modeling breast cancer in general and hormone-sensitive breast cancer, in particular in mice, has several limitations. These are related to the inbred nature of laboratory mice, and do not allow adequate appreciation of the contribution of the hosts genetic heterogeneity in tumor growth. In addition, the naturally low estrogen levels of mice makes estradiol supplementation obligatory for tumor growth. Here, we show that Peromyscus californicus, following cyclosporine-mediated immunosuppression, supports the growth of both MDA-MB-231 estrogen-independent and MCF7 estrogen receptor-positive breast cancers without exogenous estradiol supplementation. Tumor growth was inhibited by fulvestrant or letrozole, confirming that MCF7 xenografts remain hormone dependent in vivo and suggesting that P. californicus can be used as an alternative to conventional mice for the study of hormone-sensitive breast cancer. The fact that Peromyscus stocks are outbred also facilitates the study of breast cancer in genetically heterogenous populations. Summary: Outbred stocks of Peromyscus californicus (California mice), upon pharmacological immunosuppression, provide an alternative to conventional inbred mice models, and can support the growth of hormone-insensitive and hormone-sensitive human breast cancers.

Peromyscus as a model of human disease

Animals of the genus Peromyscus have been a particularly informative model for many areas of study, including behavior, evolution, anatomy, physiology and genetics. While their use in modeling human disease and pathology has been relatively restricted, certain qualities of Peromyscine mice may make them a good candidate for such studies. Pathophysiological conditions where Peromyscus may be of particular value involve aging, reactive oxygen species-associated pathologies, metabolism and detoxification, diabetes, and certain cancers. In this review article we will summarize pathological conditions where Peromyscus have been used effectively, we will discuss factors limiting the use of Peromyscus in studying pathology and we will indicate areas at which the use of this model may be of special value.