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Houston, TX, United States

Szczesny B.,University of Texas Medical Branch | Brunyanszki A.,University of Texas Medical Branch | Olah G.,University of Texas Medical Branch | Mitra S.,Houston Methodist Research Institute | Szabo C.,University of Texas Medical Branch
Nucleic Acids Research

The positive role of PARP1 in regulation of various nuclear DNA transactions is well established. Although a mitochondrial localization of PARP1 has been suggested, its role in the maintenance of the mitochondrial DNA is currently unknown. Here we investigated the role of PARP1 in the repair of the mitochondrial DNA in the baseline and oxidative stress conditions. We used wild-type A549 cells or cells depleted of PARP1. Our data show that intra-mitochondrial PARP1 interacts with a key mitochondrial-specific DNA base excision repair (BER) enzymes, namely EXOG and DNA polymerase gamma (Polγ), which under oxidative stress become poly(ADP-ribose)lated (PARylated). Interaction between mitochondrial BER enzymes was significantly affected in the presence of PARP1. Moreover, the repair of the oxidative-induced damage to the mitochondrial DNA in PARP1-depleted cells was found to be more robust compared to control counterpart. In addition, mitochondrial biogenesis was enhanced in PARP1-depleted cells, including mitochondrial DNA copy number and mitochondrial membrane potential. This observation was further confirmed by analysis of lung tissue isolated from WT and PARP1 KO mice. In summary, we conclude that mitochondrial PARP1, in opposite to nuclear PARP1, exerts a negative effect on several mitochondrial-specific transactions including the repair of the mitochondrial DNA. © 2014 The Author(s). Source

Wang F.,Tongji University | Yang Y.,Houston Methodist Research Institute
Breast Cancer Research and Treatment

The xCT antiporter is known to be upregulated in 30 % of triple-negative breast cancer (TNBC) cell lines. The xCT-CD44 variant (CD44v) system regulates the levels of reactive oxygen species (ROS) in cancer cells and promotes tumor growth. Here, the role of this antiporter system in relation to chemotherapy was evaluated. MDA-MB-231 and MDA-MB-436 cells were transfected with lentiviral vectors expressing short hairpin RNA against xCT or CD44v. Following doxorubicin treatment, cellular proliferation was monitored, ROS were measured, and intracellular levels of cysteine and glutathione (GSH) were determined using liquid chromatography-mass spectrometry. A TNBC orthotopic tumor model was used to evaluate the impact of xCT-CD44v inhibition on doxorubicin efficacy in vivo. Doxorubicin treatment of TNBC cells caused increased expression of xCT through upregulation of CD44v. Consequently, the intracellular uptake of cystine increased, enabling rapid synthesis of GSH, and neutralization of doxorubicin-induced ROS. Suppression of xCT or CD44v impaired the defense against drug-induced oxidative stress, thereby sensitizing cells to doxorubicin. The importance of the xCT-CD44v in supporting tumor growth during doxorubicin treatment was also demonstrated in an in vivo tumor model of TNBC. These findings suggest that the antiporter system could serve as a target for increasing the anticancer efficacy of conventional therapy in patients with TNBC. © 2014 Springer Science+Business Media. Source

Thomas A.,Clinical Pharmacy Services | Peterson L.E.,Houston Methodist Research Institute
International Journal of Nephrology and Renovascular Disease

Background: Ferric citrate is a novel phosphate binder which has the potential to reduce usage of erythropoietin-stimulating agents (ESAs) and intravenous (IV) iron used for anemia management during hemodialysis (HD) among patients with end-stage renal disease (ESRD). Currently, the potential health care cost savings on a national scale due to the use of ferric citrate in ESRD are undetermined. Methods: Per-patient-per-year costs of ESAs (Epogen® and Aranesp® [Amgen Inc., CA, USA]) and IV iron (Venofer® [American Regent, Inc., NY, USA] and Ferrlecit® [Sanofi US, Bridgewater, NJ, USA]) were based on RED BOOK™ (Truven Health Analytics New York, NY, USA) costs combined with the Centers for Medicare and Medicaid Services (CMS) base rate and actual usage in 2011 for the four drugs. The annual number of outpatients undergoing HD in the US was based on frequencies reported by the USRDS (United States Renal Data System). Monte Carlo uncertainty analysis was performed to determine total annual costs and cost reduction based on ferric citrate usage. Results: Total annual cost of ESAs and IV iron for anemia management in ESRD determined by Monte Carlo analysis assuming CMS base rate value was 5.127 (3.664-6.260) billion USD. For actual utilization in 2011, total annual cost of ESAs and IV iron was 3.981 (2.780-4.930) billion USD. If ferric citrate usage reduced ESA utilization by 20% and IV iron by 40%, then total cost would be reduced by 21.2% to 4.038 (2.868-4.914) billion USD for the CMS base rate, and by 21.8% to 3.111 (2.148-3.845) billion USD, based on 2011 actual utilization. Conclusion: It is likely that US health care costs for anemia-management drugs associated with ESRD among HD patients can be reduced by using ferric citrate as a phosphate binder. © 2014 Thomas and Peterson. Source

Bayrer J.R.,University of California at San Francisco | Mukkamala S.,University of California at San Francisco | Sablin E.P.,University of California at San Francisco | Webb P.,Houston Methodist Research Institute | Fletterick R.J.,University of California at San Francisco
Proceedings of the National Academy of Sciences of the United States of America

Colorectal cancers (CRCs) account for nearly 10% of all cancer deaths in industrialized countries. Recent evidence points to a central role for the nuclear receptor liver receptor homolog-1 (LRH-1) in intestinal tumorigenesis. Interaction of LRH-1 with the Wnt/β-catenin pathway, highly active in a critical subpopulation of CRC cells, underscores the importance of elucidating LRH-1's role in this disease. Reduction of LRH-1 diminishes tumor burden in murine models of CRC; however, it is not known whether LRH-1 is required for tumorigenesis, for proliferation, or for both. In this work, we address this question through shRNA-mediated silencing of LRH-1 in established CRC cell lines. LRH-1 mRNA knockdown results in significantly impaired proliferation in a cell line highly expressing the receptor and more modest impairment in a cell line with moderate LRH-1 expression. Cell-cycle analysis shows prolongation of G0/G1 with LRH-1 silencing, consistent with LRH-1 cellcycle influences in other tissues. Cluster analysis of microarray gene expression demonstrates significant genome wide alterations with major effects in cell-cycle regulation, signal transduction, bile acid and cholesterol metabolism, and control of apoptosis. This study demonstrates a critical proproliferative role for LRH-1 in established colon cancer cell lines. LRH-1 exerts its effects via multiple signaling networks. Our results suggest that selected CRC patients could benefit from LRH-1 inhibitors. Source

Chong A.S.,University of Chicago | Sciammas R.,Houston Methodist Research Institute

Much of the research on the humoral response to allografts has focused on circulating serum antibodies and the long-lived plasma cells that produce these antibodies. In contrast, the interrogation of the quiescent memory B cell compartment is technically more challenging and thus has not been incorporated into the clinical diagnostic or prognostic toolkit. In this review, we discuss new technologies that have allowed this heretofore enigmatic subset of B cells to be identified at quiescence and during a recall response. These technologies in experimental models are providing new insights into memory B cell heterogeneity with respect to their phenotype, cellular function, and the antibodies they produce. Similar technologies are also allowing for the identification of comparable memory alloreactive B cells in transplant recipients. Although much of the focus in transplant immunology has been on controlling the alloreactive B cell population, long-term transplant patient survival is also critically dependent on protection by pathogen-specific memory B cells. Techniques are available that allow the interrogation of memory B cell response to pathogen re-encounter. Thus, we are poised in our ability to investigate how immunosuppression affects allospecific and pathogen-specific memory B cells, and reason that these investigations can yield new insights that will be beneficial for graft and patient survival. Copyright © 2014 Wolters Kluwer Health, Inc. All rights reserved. Source

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