Regulation of aromatase induction by nuclear receptor coregulator PELP1

Estradiol (E2), estrogen receptor (ER), ER-coregulators have been implicated in the development and progression of breast cancer. In situ E2 synthesis is implicated in tumor cell proliferation through autocrine or paracrine mechanisms, especially in post-menopausal women. Several recent studies demonstrated activity of aromatase P450 (Cyp19), a key enzyme that plays critical role in E2 synthesis in breast tumors. The mechanism by which tumors enhance aromatase expression is not completely understood. Recent studies from our laboratory suggested that PELP1 (Proline, Glutamic acid, Leucine rich Protein 1), a novel ER-coregulator, functions as a potential proto-oncogene and promotes tumor growth in nude mice models without exogenous E2 supplementation. In this study, we found that PELP1 deregulation contributes to increased expression of aromatase, local E2 synthesis and PELP1 cooperates with growth factor signaling components in the activation of aromatase. PELP1 deregulation uniquely up-regulated aromatase expression via activation of aromatase promoter I.3/II. Analysis of PELP1 driven mammary tumors in xenograft as well as in transgenic mouse models revealed increased aromatase expression. PELP1-mediated induction of aromatase requires functional Src and PI3K pathways. Chromatin immuno precipitation (ChIP) assays revealed that PELP1 is recruited to the Aro 1.3/II aromatase promoter. HER2 signaling enhances PELP1 recruitment to the aromatase promoter and PELP1 plays a critical role in HER2-mediated induction of aromatase expression. Mechanistic studies revealed that PELP1 interactions with orphan receptor ERRα, and histone demethylases play a role in the activation of aromatase promoter. Accordingly, ChIP analysis showed alterations in histone modifications at the aromatase promoter in the model cells that exhibit local E2 synthesis. Immunohistochemical analysis of breast tumor progression tissue arrays suggested that deregulation of aromatase expression occurs in advanced-stage and node-positive tumors, and that cooverexpression of PELP1 and aromatase occur in a sub set of tumors. Collectively, our results suggest that PELP1 regulation of aromatase represent a novel mechanism for in situ estrogen synthesis leading to tumor proliferation by autocrine loop and open a new avenue for ablating local aromatase activity in breast tumors.     

Tumour escape mechanisms and their therapeutic implications in combination tumour therapy

Most tumours arise from a single normal cell through a sequential evolutionary process of mutation and selection. Tumours are initiated by escaping non‐immune surveillance, which includes defective DNA repair, epigenetic gene alternation, resistance to apoptosis and loss of intercellular contact inhibition. Tumour cells harbour mutations in a number of critical genes that provide selective advantages at various stages during the evolution of the tumour. The tumour cells that circumvent the tumour suppressor mechanisms of the non‐immune surveillance process are edited by the immune system, resulting in the selection of a resistant tumour variant. The selection of the tumour cell is further shaped by its interactions with cells and other factors in its microenvironment. Tumour evolution is thought to adhere to Darwinian principles by escaping both non‐immune (intrinsic) and immune (extrinsic) responses against self‐altered tumour cells. At end‐stage, tumours have escaped both non‐immune and immune surveillance with increased threshold of apoptosis. Combination therapy has been proposed, by exploring the non‐immune and immune suppressive nature of the tumour, and has been found to have a therapeutic efficiency on tumour regression as compared with monotherapies. The combination of immunotherapy and other different modalities, especially vaccines, with conventional anticancer therapies with optimized dosage and scheduling can offer synergistic antitumour effects. Here, we focus on the mechanism of tumour evolution and its implication in combination therapy.     

G-protein activation modulates pseudo-periodic oscillation of Na channel

We have shown before that the duration and amplitude of both prolonged (1-160 s) and short (100-1000 ms) depolarizing prepulse altered all the steady-state and kinetic parameters of rNav1.2a voltage-gated sodium channel in a pseudo-oscillatory fashion with variable time period and amplitude, often superimposed on a linear trend. In this study, we have examined the effect of G-protein activation on pseudo-oscillatory properties of the rNav1.2a sodium channel alpha subunit, heterologously expressed in Chinese hamster ovary cells. G-protein modification caused insignificant changes in the slow pseudo-periodic oscillation of the activation properties of sodium channel; only the time period of the oscillation was altered from approximately 30 to 21s. In contrast, G-protein activation abolished the faster component of pseudo-periodic oscillation in steady-state inactivation properties of sodium channel; the conditioning duration dependence of steady-state inactivation becomes monotonic in nature.     

Kynurenate treatment of autaptic hippocampal microcultures affect localized voltage-dependent calcium diffusion in the dendrites

It is not clear how different spatial compartments in the neuron are affected during epileptiform activity. In the present study we have examined the spatial and temporal profiles of depolarization induced changes in the intracellular Ca(2+) concentration in the dendrites of cultured autaptic hippocampal pyramidal neurons rendered epileptic experimentally by treatment with kynurenate (2 mM) and Mg(2+) (11.3 mM) in culture (treated neurons). This was examined with simultaneous somatic patch-pipette recording and Ca(2+) imaging experiments using the Ca(2+) indicator Oregon Green 488 BAPTA-1. Neurons stimulated by depolarization under whole-cell voltage clamp conditions revealed Ca(2+) entry at localized sites in the dendrites. Ca(2+) transients were observed even in the presence of NMDA and AMPA receptor antagonists suggesting that the opening of voltage gated calcium channels primarily triggered the local Ca(2+) changes. Peak Ca(2+) transients in the dendrites of treated neurons were larger compared to the signals recorded from the control neurons. Dendritic Ca(2+) transients in treated neurons showed a distance dependent scaling. Estimation of dendritic local Ca(2+) diffusion coefficients indicated higher values in the treated neurons and a higher availability of free Ca(2+). Simulation studies of Ca(2+) dynamics in these localized dendritic compartments indicate that local Ca(2+) buffering and removal mechanisms may be affected in treated neurons. Our studies indicate that small dendritic compartments are rendered more vulnerable to changes in intracellular Ca(2+) following induction of epileptiform activity. This can have important cellular consequences including local membrane excitability through mechanisms that remain to be elucidated.     

Spatial association between speciated fine particles and mortality

Particulate matter (PM) has been linked to a range of serious cardiovascular and respiratory health problems, including premature mortality. The main objective of our research is to quantify uncertainties about the impacts of fine PM exposure on mortality. We develop a multivariate spatial regression model for the estimation of the risk of mortality associated with fine PM and its components across all counties in the conterminous United States. We characterize different sources of uncertainty in the data and model the spatial structure of the mortality data and the speciated fine PM. We consider a flexible Bayesian hierarchical model for a space-time series of counts (mortality) by constructing a likelihood-based version of a generalized Poisson regression model that combines methods for point-level misaligned data and change of support regression. Our results seem to suggest an increase by a factor of two in the risk of mortality due to fine particles with respect to coarse particles. Our study also shows that in the Western United States, the nitrate and crustal components of the speciated fine PM seem to have more impact on mortality than the other components. On the other hand, in the Eastern United States, sulfate and ammonium explain most of the fine PM effect.     

Antiatherogenic and radioprotective role of folic acid in whole body γ-irradiated mice

Free radical mediated oxidative damage is one of the prime factors for atherogenic changes in humans. We have shown that the folic acid administration reduced the risk of the atherogenic factors induced by γ -radiation. Folic acid administration prevented the radiation induced increase in the plasma lipoprotein lipase activity and also prevented the radiation-induced increase in the hepatic cholesterol and triglycerides levels. These results indicate the role of folic acid as an antiatherogenic agent. Further, we also report the radioprotective property of folic acid as demonstrated by reduction in the radiation induced membrane damage as measured by lipid peroxidation products and DNA damage, which was measured by alkaline comet assay.     

Preferential migration of regulatory T cells mediated by gliomasecreted chemokines can be blocked with chemotherapy

Despite the immunogenicity of glioblastoma multiforme (GBM), immune-mediated eradication of these tumors remains deficient. Regulatory T cells (Tregs) in the blood and within the tumor microenvironment of GBM patients are known to contribute to their dismal immune responses. Here, we determined which chemokine secreted by gliomas can preferentially induce Treg recruitment and migration. In the malignant human glioma cell lines D-54, U-87, U-251, and LN-229, the chemokines CCL22 and CCL2 were detected by intracellular cytokine analysis. Furthermore, tumor cells from eight patients with GBM had a similar chemokine expression profile. However, only CCL2 was detected by enzyme-linked immunosorbent assay, indicating that CCL2 may be the principal chemokine for Treg migration in GBM patients. Interestingly, the Tregs from GBM patients had significantly higher expression levels of the CCL2 receptor CCR4 than did Tregs from healthy controls. Glioma supernatants and the recombinant human chemokines CCL2 and CCL22 induced Treg migration and were blocked by antibodies to the chemokine receptors. Production of CCL2 by glioma cells could also be mitigated by the chemotherapeutic agents temozolomide and carmustine [3-bis (2-chloroethyl)-1-nitrosourea]. Our results indicate that gliomas augment immunosuppression by selective chemokine-mediated recruitment of Tregs into the tumor microenvironment and that modulating this interaction with chemotherapy could facilitate the development of novel immunotherapeutics to malignant gliomas. Justin T. Jordan and Wei Sun are contributed equally to this work. An erratum to this article can be found at