ER 50891

ZEB1 induces ER-alpha promoter hypermethylation and confers antiestrogen resistance in breast cancer.[Pubmed:28383555]

Cell Death Dis. 2017 Apr 6;8(4):e2732.

Antiestrogen resistance is a major obstacle to endocrine therapy for breast cancers. Although reduced estrogen receptor-alpha (ER-alpha) expression is a known contributing factor to antiestrogen resistance, the mechanisms of ER-alpha downregulation in antiestrogen resistance are not fully understood. Here, we report that ectopic zinc-finger E-box binding homeobox 1 (ZEB1) is associated with ER-alpha deficiency in breast cancer cells and thus confers antiestrogen resistance. Mechanistically, ZEB1 represses ER-alpha transcription by forming a ZEB1/DNA methyltransferase (DNMT)3B/histone deacetylase (HDAC)1 complex on the ER-alpha promoter, leading to DNA hypermethylation and the silencing of ER-alpha. Thus, ectopic ZEB1 downregulates ER-alpha expression and subsequently attenuates cell growth inhibition by antiestrogens, such as tamoxifen and fulvestrant. Notably, the depletion of ZEB1 by RNA interference causes ER-alpha promoter demethylation, restores ER-alpha expression, and increases the responsiveness of breast cancer cells to antiestrogen treatment. By studying specimens from a large cohort of subjects with breast cancer, we found a strong inverse correlation between ZEB1 and ER-alpha protein expression. Moreover, breast tumors that highly express ZEB1 exhibit ER-alpha promoter hypermethylation. Using a nude mouse xenograft model, we further confirmed that the downregulation of ZEB1 expression restores the responsiveness of breast cancer cells to antiestrogen therapy in vivo. Therefore, our findings suggest that ZEB1 is a crucial determinant of resistance to antiestrogen therapies in breast cancer.

GSK-3-mediated phosphorylation couples ER-Golgi transport and nuclear stabilization of the CREB-H transcription factor to mediate apolipoprotein secretion.[Pubmed:28381424]

Mol Biol Cell. 2017 Jun 1;28(11):1565-1579.

CREB-H, an ER-anchored transcription factor, plays a key role in regulating secretion in metabolic pathways, particularly triglyceride homeostasis. It controls the production both of secretory pathway components and cargoes, including apolipoproteins ApoA-IV and ApoC-II, contributing to VLDL/HDL distribution and lipolysis. The key mechanism controlling CREB-H activity involves its ER retention and forward transport to the Golgi, where it is cleaved by Golgi-resident proteases, releasing the N-terminal product, which traffics to the nucleus to effect transcriptional responses. Here we show that a serine-rich motif termed the P-motif, located in the N-terminus between serines 73 and 90, controls release of the precursor transmembrane form from the ER and its forward transport to the Golgi. This motif is subject to GSK-3 phosphorylation, promoting ER retention, while mutation of target serines and drug inhibition of GSK-3 activity coordinately induce both forward transport of the precursor and cleavage, resulting in nuclear import. We previously showed that for the nuclear product, the P-motif is subject to multiple phosphorylations, which regulate stability by targeting the protein to the SCF(Fbw1a) E3 ubiquitin ligase. Thus phosphorylation at the P-motif provides integrated control of CREB-H function, coupling intercompartmental transport in the cytoplasm with stabilization of the active form in the nucleus.

Physiochemical properties of greatly enhanced photoluminescence of aqueous dispersible upconversion CaF2:Yb/Er nanoparticles.[Pubmed:28382337]

Photochem Photobiol Sci. 2017 Jun 14;16(6):890-896.

Crystal phase morphological structure and optical properties of the as-prepared upconversion CaF2:Yb/Er(core) and sequential coating of an inert crystalline material and silica layers surrounding the seed core-nanoparticles (NPs) were characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), selected area electron diffraction (SAED), optical absorption, FTIR spectroscopy and upconversion photoluminescence spectroscopy. Owing to the unique properties of CaF2 host matrix, we realized their practical applications in biomedical science to improve the upconversion luminescence property and aqueous dispersibility. The surface coating on the seed core particles will significantly influence the structural, optical band gap energy and upconversion luminescence properties. These NPs were well-dispersed in aqueous and non-aqueous solvents to form clear colloidal solutions. The colloidal solutions of three samples show a characteristic optical absorption band in UV/Visible region. As a result, optical band gap gradually decreases after sequential growth of the inert shell and amorphous silica due to an increase in the crystalline size. Comparative upconversion luminescence analysis showed that after inert shell growth, the upconversion intensity was greatly improved, and such an improvement was found to arise from efficient suppression of surface-related deactivation from the core nanocrystals. Interestingly, growth of an inert (CaF2) shell over the seed core NPs shows intense upconversion emission lines under 980 nm NIR laser excitation, highlighting their promising applications, such as multi-analyte biolabels, staining, displays and other photonic based technological applications.

PMP22 exon 4 deletion causes ER retention of PMP22 and a gain-of-function allele in CMT1E.[Pubmed:28382305]

Ann Clin Transl Neurol. 2017 Mar 12;4(4):236-245.

OBJECTIVE: To determine whether predicted fork stalling and template switching (FoSTeS) during mitosis deletes exon 4 in peripheral myelin protein 22 KD (PMP22) and causes gain-of-function mutation associated with peripheral neuropathy in a family with Charcot-Marie-Tooth disease type 1E. METHODS: Two siblings previously reported to have genomic rearrangements predicted to involve exon 4 of PMP22 were evaluated clinically and by electrophysiology. Skin biopsies from the proband were studied by RT-PCR to determine the effects of the exon 4 rearrangements on exon 4 mRNA expression in myelinating Schwann cells. Transient transfection studies with wild-type and mutant PMP22 were performed in Cos7 and RT4 cells to determine the fate of the resultant mutant protein. RESULTS: Both affected siblings had a sensorimotor dysmyelinating neuropathy with severely slow nerve conduction velocities (<10 m/sec). RT-PCR studies of Schwann cell RNA from one of the siblings demonstrated a complete in-frame deletion of PMP22 exon 4 (PMP22Delta4). Transfection studies demonstrated that PMP22Delta4 protein is retained within the endoplasmic reticulum and not transported to the plasma membrane. CONCLUSIONS: Our results confirm that that FoSTeS-mediated genomic rearrangement produced a deletion of exon 4 of PMP22, resulting in expression of both PMP22 mRNA and protein lacking this sequence. In addition, we provide experimental evidence for endoplasmic reticulum retention of the mutant protein suggesting a gain-of-function mutational mechanism consistent with the observed CMT1E in this family. PMP22Delta4 is another example of a mutated myelin protein that is misfolded and contributes to the pathogenesis of the neuropathy.

Disruption of retinoic acid receptor alpha reveals the growth promoter face of retinoic acid.[Pubmed:17786207]

PLoS One. 2007 Sep 5;2(9):e836.

BACKGROUND: Retinoic acid (RA), the bioactive derivative of Vitamin A, by epigenetically controlling transcription through the RA-receptors (RARs), exerts a potent antiproliferative effect on human cells. However, a number of studies show that RA can also promote cell survival and growth. In the course of one of our studies we observed that disruption of RA-receptor alpha, RARalpha, abrogates the RA-mediated growth-inhibitory effects and unmasks the growth-promoting face of RA (Ren et al., Mol. Cell. Biol., 2005, 25:10591). The objective of this study was to investigate whether RA can differentially govern cell growth, in the presence and absence of RARalpha, through differential regulation of the "rheostat" comprising ceramide (CER), the sphingolipid with growth-inhibitory activity, and sphingosine-1-phosphate (S1P), the sphingolipid with prosurvival activity. METHODOLOGY/PRINCIPAL FINDINGS: We found that functional inhibition of endogenous RARalpha in breast cancer cells by using either RARalpha specific antagonists or a dominant negative RARalpha mutant hampers on one hand the RA-induced upregulation of neutral sphingomyelinase (nSMase)-mediated CER synthesis, and on the other hand the RA-induced downregulation of sphingosine kinase 1, SK1, pivotal for S1P synthesis. In association with RA inability to regulate the sphingolipid rheostat, cells not only survive, but also grow more in response to RA both in vitro and in vivo. By combining genetic, pharmacological and biochemical approaches, we mechanistically demonstrated that RA-induced growth is, at least in part, due to non-RAR-mediated activation of the SK1-S1P signaling. CONCLUSIONS/SIGNIFICANCE: In the presence of functional RARalpha, RA inhibits cell growth by concertedly, and inversely, modulating the CER and S1P synthetic pathways. In the absence of a functional RARalpha, RA-in a non-RAR-mediated fashion-promotes cell growth by activating the prosurvival S1P signaling. These two distinct, yet integrated processes apparently concur to the growth-promoter effects of RA.

Impaired retinoic acid (RA) signal leads to RARbeta2 epigenetic silencing and RA resistance.[Pubmed:16287870]

Mol Cell Biol. 2005 Dec;25(23):10591-603.

Resistance to the growth-inhibitory action of retinoic acid (RA), the bioactive derivative of vitamin A, is common in human tumors. One form of RA resistance has been associated with silencing and hypermethylation of the retinoic acid receptor beta2 gene (RARbeta2), an RA-regulated tumor suppressor gene. The presence of an epigenetically silent RARbeta2 correlates with lack of the RA receptor alpha (RARalpha). Normally, RARalpha regulates RARbeta2 transcription by mediating dynamic changes of RARbeta2 chromatin in the presence and absence of RA. Here we show that interfering with RA signal through RARalpha (which was achieved by use of a dominant-negative RARalpha, by downregulation of RARalpha by RNA interference, and by use of RARalpha antagonists) induces an exacerbation of the repressed chromatin status of RARbeta2 and leads to RARbeta2 transcriptional silencing. Further, we demonstrate that RARbeta2 silencing causes resistance to the growth-inhibitory effect of RA. Apparently, RARbeta2 silencing can also occur in the absence of DNA methylation. Conversely, we demonstrate that restoration of RA signal at a silent RARbeta2 through RARalpha leads to RARbeta2 reactivation. This report provides proof of principle that RARbeta2 silencing and RA resistance are consequent to an impaired integration of RA signal at RARbeta2 chromatin.

Syntheses and evaluation of quinoline derivatives as novel retinoic acid receptor alpha antagonists.[Pubmed:11354380]

Bioorg Med Chem Lett. 2001 May 7;11(9):1215-8.

In the course of studies on novel retinoids, we have designed and synthesized a series of quinoline derivatives. One of them, 4-[5-[8-(1-methylethyl)-4-phenyl-2-quinolinyl]-1H-2-pyrrolyl]benzoic acid (12f) shows potent RARalpha-selective antagonistic activity.