Targeting GSK3 and Associated Signaling Pathways Involved in Cancer

Glycogen synthase kinase 3 (GSK-3) is a serine/threonine (S/T) protein kinase. Although GSK-3 originally was identified to have functions in regulation of glycogen synthase, it was subsequently determined to have roles in multiple normal biochemical processes as well as various disease conditions. GSK-3 is sometimes referred to as a moonlighting protein due to the multiple substrates and processes which it controls. Frequently, when GSK-3 phosphorylates proteins, they are targeted for degradation. GSK-3 is often considered a component of the PI3K/PTEN/AKT/GSK-3/mTORC1 pathway as GSK-3 is frequently phosphorylated by AKT which regulates its inactivation. AKT is often active in human cancer and hence, GSK-3 is often inactivated. Moreover, GSK-3 also interacts with WNT/\u3b2-catenin signaling and \u3b2-catenin and other proteins in this pathway are targets of GSK-3. GSK-3 can modify NF-\u3baB activity which is often expressed at high levels in cancer cells. Multiple pharmaceutical companies developed small molecule inhibitors to suppress GSK-3 activity. In addition, various natural products will modify GSK-3 activity. This review will focus on the effects of small molecule inhibitors and natural products on GSK-3 activity and provide examples where these compounds were effective in suppressing cancer growth

Biochar versus hydrochar as growth media constituents for ornamental plant cultivation

[EN] Biochar and hydrochar have been proposed as novel materials for providing soilless growth media. However, much more knowledge is required before reliable advice can be given on the use of these materials for this purpose. Depending on the material and the technology applied (pyrolysis or hydrothermal carbonization), phytotoxicity and greenhouse gas emissions have been found for certain chars. In this study, our aim was to assess the feasibility of three chars as substrate constituents. We compared two biochars, one from forest waste and the other from olive mill waste, and a hydrochar from forest waste. We studied how chars affected substrate characteristics, plant performance, water economy and respiratory CO2 emission. Substrates containing biochar from forest waste showed the best characteristics, with good air/water relationships and adequate electrical conductivity. Those with biochar from olive mill waste were highly saline and, consequently, low quality. The substrates with hydrochar retained too much water and were poorly aerated, presenting high CO2 concentrations due to high respiratory activity. Plants performed well only when grown in substrates containing a maximum of 25 % biochar from forest waste or hydrochar. After analyzing the char characteristics, we concluded that biochar from forest waste could be safely used as a substrate constituent and is environmentally friendly when applied due to its low salinity and low CO2 emission. However, biochar from olive mill waste and hydrochar need to be improved before they can be used as substrate constituents.This study was funded by the Polytechnic University of Valencia (Projects on New Multidisciplinary Research; PAID-05-12). We thank Molly Marcus-McBride for supervising the English.Fornes Sebastiá, F.; Belda Navarro, RM. (2018). Biochar versus hydrochar as growth media constituents for ornamental plant cultivation. Scientia Agricola (Online). 75(4):304-312. https://doi.org/10.1590/1678-992X-2017-0062S304312754Abad, M., Noguera, P., & Burés, S. (2001). National inventory of organic wastes for use as growing media for ornamental potted plant production: case study in Spain. Bioresource Technology, 77(2), 197-200. doi:10.1016/s0960-8524(00)00152-8Bargmann, I., Martens, R., Rillig, M. C., Kruse, A., & Kücke, M. (2013). Hydrochar amendment promotes microbial immobilization of mineral nitrogen. Journal of Plant Nutrition and Soil Science, 177(1), 59-67. doi:10.1002/jpln.201300154Bargmann, I., Rillig, M. C., Buss, W., Kruse, A., & Kuecke, M. (2013). Hydrochar and Biochar Effects on Germination of Spring Barley. Journal of Agronomy and Crop Science, 199(5), 360-373. doi:10.1111/jac.12024Bedussi, F., Zaccheo, P., & Crippa, L. (2015). Pattern of pore water nutrients in planted and non-planted soilless substrates as affected by the addition of biochars from wood gasification. Biology and Fertility of Soils, 51(5), 625-635. doi:10.1007/s00374-015-1011-6Belda, R. M., Lidón, A., & Fornes, F. (2016). Biochars and hydrochars as substrate constituents for soilless growth of myrtle and mastic. Industrial Crops and Products, 94, 132-142. doi:10.1016/j.indcrop.2016.08.024Costello, R. C., & Sullivan, D. M. (2013). Determining the pH Buffering Capacity of Compost Via Titration with Dilute Sulfuric Acid. Waste and Biomass Valorization, 5(3), 505-513. doi:10.1007/s12649-013-9279-yFernandes, C., & Corá, J. E. (2004). Bulk density and relationship air/water of horticultural substrate. Scientia Agricola, 61(4), 446-450. doi:10.1590/s0103-90162004000400015Fornes, F., Belda, R. M., Carrión, C., Noguera, V., García-Agustín, P., & Abad, M. (2007). Pre-conditioning ornamental plants to drought by means of saline water irrigation as related to salinity tolerance. Scientia Horticulturae, 113(1), 52-59. doi:10.1016/j.scienta.2007.01.008Fornes, F., Belda, R. M., & Lidón, A. (2015). Analysis of two biochars and one hydrochar from different feedstock: focus set on environmental, nutritional and horticultural considerations. Journal of Cleaner Production, 86, 40-48. doi:10.1016/j.jclepro.2014.08.057Fornes, F., & Belda, R. M. (2017). Acidification with nitric acid improves chemical characteristics and reduces phytotoxicity of alkaline chars. Journal of Environmental Management, 191, 237-243. doi:10.1016/j.jenvman.2017.01.026Fornes, F., Belda, R. M., Fernández de Córdova, P., & Cebolla-Cornejo, J. (2017). Assessment of biochar and hydrochar as minor to major constituents of growing media for containerized tomato production. Journal of the Science of Food and Agriculture, 97(11), 3675-3684. doi:10.1002/jsfa.8227Fornes, F., Carrión, C., García-de-la-Fuente, R., Puchades, R., & Abad, M. (2010). Leaching composted lignocellulosic wastes to prepare container media: Feasibility and environmental concerns. Journal of Environmental Management, 91(8), 1747-1755. doi:10.1016/j.jenvman.2010.03.017GARCIADELAFUENTE, R., CARRION, C., BOTELLA, S., FORNES, F., NOGUERA, V., & ABAD, M. (2007). Biological oxidation of elemental sulphur added to three composts from different feedstocks to reduce their pH for horticultural purposes. Bioresource Technology, 98(18), 3561-3569. doi:10.1016/j.biortech.2006.11.008Genty, B., Briantais, J.-M., & Baker, N. R. (1989). The relationship between the quantum yield of photosynthetic electron transport and quenching of chlorophyll fluorescence. Biochimica et Biophysica Acta (BBA) - General Subjects, 990(1), 87-92. doi:10.1016/s0304-4165(89)80016-9Hoitink, H. A. J., Stone, A. G., & Han, D. Y. (1997). Suppression of Plant Diseases by Composts. HortScience, 32(2), 184-187. doi:10.21273/hortsci.32.2.184Libra, J. A., Ro, K. S., Kammann, C., Funke, A., Berge, N. D., Neubauer, Y., … Emmerich, K.-H. (2011). Hydrothermal carbonization of biomass residuals: a comparative review of the chemistry, processes and applications of wet and dry pyrolysis. Biofuels, 2(1), 71-106. doi:10.4155/bfs.10.81Mazuela, P., Salas, M. del C., & Urrestarazu, M. (2005). Vegetable Waste Compost as Substrate for Melon. Communications in Soil Science and Plant Analysis, 36(11-12), 1557-1572. doi:10.1081/css-200059054Méndez, A., Paz-Ferreiro, J., Gil, E., & Gascó, G. (2015). The effect of paper sludge and biochar addition on brown peat and coir based growing media properties. Scientia Horticulturae, 193, 225-230. doi:10.1016/j.scienta.2015.07.032Nieto, A., Gascó, G., Paz-Ferreiro, J., Fernández, J. M., Plaza, C., & Méndez, A. (2016). The effect of pruning waste and biochar addition on brown peat based growing media properties. Scientia Horticulturae, 199, 142-148. doi:10.1016/j.scienta.2015.12.012Sáez, J. A., Belda, R. M., Bernal, M. P., & Fornes, F. (2016). Biochar improves agro-environmental aspects of pig slurry compost as a substrate for crops with energy and remediation uses. Industrial Crops and Products, 94, 97-106. doi:10.1016/j.indcrop.2016.08.035Smith, B. R., Fisher, P. R., & Argo, W. R. (2004). Growth and Pigment Content of Container-grown Impatiens and Petunia in Relation to Root Substrate pH and Applied Micronutrient Concentration. HortScience, 39(6), 1421-1425. doi:10.21273/hortsci.39.6.1421Solaiman, Z. M., Murphy, D. V., & Abbott, L. K. (2011). Biochars influence seed germination and early growth of seedlings. Plant and Soil, 353(1-2), 273-287. doi:10.1007/s11104-011-1031-4Steiner, C., & Harttung, T. (2014). Biochar as a growing media additive and peat substitute. Solid Earth, 5(2), 995-999. doi:10.5194/se-5-995-2014Tian, Y., Sun, X., Li, S., Wang, H., Wang, L., Cao, J., & Zhang, L. (2012). Biochar made from green waste as peat substitute in growth media for Calathea rotundifola cv. Fasciata. Scientia Horticulturae, 143, 15-18. doi:10.1016/j.scienta.2012.05.018Vaughn, S. F., Eller, F. J., Evangelista, R. L., Moser, B. R., Lee, E., Wagner, R. E., & Peterson, S. C. (2015). Evaluation of biochar-anaerobic potato digestate mixtures as renewable components of horticultural potting media. Industrial Crops and Products, 65, 467-471. doi:10.1016/j.indcrop.2014.10.04

The therapeutic potential of mTOR inhibitors in breast cancer.

Rapamycin and modified rapamycins (rapalogs) have been used to prevent allograft rejection after organ transplant for over 15 years. The mechanistic target of rapamycin (mTOR) has been determined to be a key component of the mTORC1 complex which consists of the serine/threonine kinase TOR and at least five other proteins which are involved in regulating its activity. Some of the best characterized substrates of mTORC1 are proteins which are key kinases involved in the regulation of cell growth (e.g., p70S6K) and protein translation (e.g., 4E-BP1). These proteins may in some cases serve as indicators to sensitivity to rapamycin-related therapies. Dysregulation of mTORC1 activity frequently occurs due to mutations at, or amplifications of, upstream growth factor receptors (e.g., human epidermal growth factor receptor-2, HER2) as well as kinases (e.g., PI3K) and phosphatases (e.g., PTEN) critical in the regulation of cell growth. More recently, it has been shown that certain rapalogs may enhance the effectiveness of hormonal-based therapies for breast cancer patients who have become resistant to endocrine therapy. The combined treatment of certain rapalogs (e.g., everolimus) and aromatase inhibitors (e.g., exemestane) has been approved by the United States Food and Drug Administration (US FDA) and other drug regulatory agencies to treat estrogen receptor positive (ER+) breast cancer patients who have become resistant to hormonal-based therapies and have progressed. This review will summarize recent basic and clinical research in the area and evaluate potential novel therapeutic approaches

Diagnostic value of neutrophil gelatinase-associated lipocalin/matrix metalloproteinase-9 pathway in transitional cell carcinoma of the bladder

Neutrophil gelatinase-associated lipocalin (NGAL), matrix metalloproteinase (MMP)-9, and NGAL/MMP-9 complex have been evaluated as diagnostic markers of several cancers, but results for bladder cancer are scanty. We evaluated these proteins in urine and serum of 89 patients with histologically confirmed bladder cancer and 119 cancer-free controls from a case-control study. Urinary concentrations were standardized on creatinine level. The performance of these proteins as cancer biomarkers was evaluated through the receiver operating characteristic (ROC) analysis. Urinary level of NGAL, MMP-9, and NGAL/MMP-9 complex was higher in current smokers, whereas no impact of dietary habits was observed. After adjusting for tobacco smoking, urinary concentration of MMP-9 was independently associated with cancer invasiveness, grading, and histological subtype, with elevated concentrations among T2\u2013T4 and non-papillary bladder cancers. Conversely, NGAL and NGAL/MMP-9 complex were significantly higher in non-papillary than in papillary subtype. The pattern was less clear in serum, but correlation between urinary and serum concentration was poor, especially for Ta/is\u2013T1 tumors. The ROC analysis confirmed that MMP-9 was the best marker (area under the ROC curve (AUC) = 0.68). Performances were much greater for muscle-invasive bladder cancers (AUC = 0.90), with elevated negative predictive values (97\ua0%). The present study suggests that NGAL/MMP-9 pathway is associated with an aggressive phenotype of bladder cancer. The elevated negative predictive value of MMP-9 and NGAL/MMP-9 complex makes them candidate markers of exclusion test for bladder cancer. These proteins may be integrated in the surveillance of bladder cancer, thus diminishing patients\u2019 discomfort and improving compliance

Enhancing therapeutic efficacy by targeting non-oncogene addicted cells with combinations of signal transduction inhibitors and chemotherapy.

The effects of inhibition of the Raf/MEK/ERK and PI3K/Akt/mTOR signaling pathways and chemotherapeutic drugs on cell cycle progression and drug sensitivity were examined in cytokine-dependent FL5.12 hematopoietic cells. We examined their effects, as these cells resemble normal hematopoietic precursor cells as they do not exhibit “oncogene-addicted” growth, while they do display “cytokine-addicted” proliferation as cytokine removal resulted in apoptosis in greater than 80% of the cells within 48 h. When cytokine-dependent FL5.12 cells were cultured in the presence of IL-3, which stimulated multiple proliferation and anti-apoptotic cascades, MEK, PI3K and mTOR inhibitors transiently suppressed but did not totally inhibit cell cycle progression or induce apoptosis while chemotherapeutic drugs such as doxorubicin and paclitaxel were more effective in inducing cell cycle arrest and apoptosis. Doxorubicin induced a G(1) block, while paclitaxel triggered a G(2)/M block. Doxorubicin was more effective in inducing cell death than paclitaxel. Furthermore the effects of doxorubicin could be enhanced by addition of MEK, PI3K or mTOR inhibitors. Cytokine-dependent cells which proliferate in vitro and are not “oncogene-addicted” may represent a pre-malignant stage, more refractory to treatment with targeted therapy. However, these cells are sensitive to chemotherapeutic drugs. It is important to develop methods to inhibit the growth of such cytokine-dependent cells as they may resemble the leukemia stem cell and other cancer initiating cells. These results demonstrate the enhanced effectiveness of targeting early hematopoietic progenitor cells with combinations of chemotherapeutic drugs and signal transduction inhibitors

Involvement of Akt-1 and mTOR in sensitivity of breast cancer to targeted therapy.

Elucidating the response of breast cancer cells to chemotherapeutic and hormonal based drugs is clearly important as these are frequently used therapeutic approaches. A signaling pathway often involved in chemo-and hormonal-resistance is the Ras/PI3K/PTEN/Akt/mTOR cascade. In the studies presented in this report, we have examined the effects of constitutive activation of Akt on the sensitivity of MCF7 breast cancer cells to chemotherapeutic-and hormonal-based drugs as well as mTOR inhibitors. MCF-7 cells which expressed a constitutively-activated Akt-1 gene [.Akt-1(CA)] were more resistant to doxorubicin, etoposide and 4-OH-tamoxifen (4HT) than cells lacking.Akt-1(CA). Cells which expressed Delta Akt-1(CA) were hypersensitive to the mTOR inhibitor rapamycin. Furthermore, rapamycin lowered the IC(50)s for doxorubicin, etoposide and 4HT in the cells which expressed Delta Akt-1(CA), demonstrating a potential improved method for treating certain breast cancers which have deregulated PI3K/PTEN/Akt/mTOR signaling. Understanding how breast cancers respond to chemo-and hormonal-based therapies and the mechanisms by which they can become drug resistant may enhance our ability to treat breast cancer. These results also document the potential importance of knowledge of the mutations present in certain cancers which may permit more effective therapies

Therapeutic resistance in breast cancer cells can result from deregulated EGFR signaling

The epidermal growth factor receptor (EGFR) interacts with various downstream molecules including phospholipase C (PLC)/protein kinase C (PKC), Ras/Raf/MEK/ERK, PI3K/PTEN/Akt/GSK-3, Jak/STAT and others. Often these pathways are deregulated in human malignancies such as breast cancer. Various therapeutic approaches to inhibit the activity of EGFR family members including small molecule inhibitors and monoclonal antibodies (MoAb) have been developed. A common problem with cancer treatments is the development of drug-resistance. We examined the effects of a conditionally-activated EGFR (v-Erb-B:ER) on the resistance of breast cancer cells to commonly used chemotherapeutic drugs such as doxorubicin, daunorubicin, paclitaxel, cisplatin and 5-flurouracil as well as ionizing radiation (IR). v-Erb-B is similar to the EGFR-variant EGFRvIII, which is expressed in various cancers including breast, brain, prostate. Both v-Erb-B and EGFRvIII encode the EGFR kinase domain but lack key components present in the extracellular domain of EGFR which normally regulate its activity and ligand-dependence. The v-Erb-B oncogene was ligated to the hormone binding domain of the estrogen receptor (ER) which results in regulation of the activity of the v-Erb-ER construct by addition of either estrogen (E2) or 4-hydroxytamoxifen (4HT) to the culture media. Introduction of the v-Erb-B:ER construct into the MCF-7 breast cancer cell line increased the resistance to the cells to various chemotherapeutic drugs, hormonal-based therapeutics and IR. These results point to the important effects that aberrant expression of EGFR kinase domain can have on therapeutic resistance