The standard low-phosphorus diet restricts pulses, nuts, and whole grains and other high phosphorus foods to control hyperphosphatemia. We conducted a randomized controlled trial to evaluate the ...effectiveness, safety, and tolerability of the modified diet, which introduced some pulses and nuts, increased the use of whole grains, increased focus on the avoidance of phosphate additives, and introduced the prescription of low-biological-value protein such as bread.
We conducted a multicenter, pragmatic, parallel-arm, open-label, randomized controlled trial of modified versus standard diet in 74 adults on hemodialysis with hyperphosphatemia over 1 month. Biochemistry was assessed using monthly laboratory tests. Dietary intake was assessed using a 2-day record of weighed intake of food, and tolerability was assessed using a patient questionnaire.
There was no significant difference in the change in serum phosphate between the standard and modified diets. Although total dietary phosphorus intake was similar, phytate-bound phosphorus, found in pulses, nuts, and whole grains, was significantly higher in the modified diet (P < 0.001). Dietary fiber intake was also significantly higher (P < 0.003), as was the percentage of patients reporting an increase in bowel movements while following the modified diet (P = 0.008). There was no significant difference in the change in serum potassium or in reported protein intake between the 2 diets. Both diets were similarly well tolerated.
The modified low phosphorus diet was well tolerated and was associated with similar phosphate and potassium control but with a wider food choice and greater fiber intake than the standard diet.
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The implementation of targeted therapies for acute myeloid leukaemia (AML) has been challenging because of the complex mutational patterns within and across patients as well as a dearth of ...pharmacologic agents for most mutational events. Here we report initial findings from the Beat AML programme on a cohort of 672 tumour specimens collected from 562 patients. We assessed these specimens using whole-exome sequencing, RNA sequencing and analyses of ex vivo drug sensitivity. Our data reveal mutational events that have not previously been detected in AML. We show that the response to drugs is associated with mutational status, including instances of drug sensitivity that are specific to combinatorial mutational events. Integration with RNA sequencing also revealed gene expression signatures, which predict a role for specific gene networks in the drug response. Collectively, we have generated a dataset-accessible through the Beat AML data viewer (Vizome)-that can be leveraged to address clinical, genomic, transcriptomic and functional analyses of the biology of AML.
Ovarian cancer is generally diagnosed at an advanced stage where the case/fatality ratio is high and thus remains the most lethal of all gynecologic malignancies among US women. Serous tumors are the ...most widespread forms of ovarian cancer and the Tg-MISIIR-TAg transgenic represents the only mouse model that spontaneously develops this type of tumors. Tg-MISIIR-TAg mice express SV40 transforming region under control of the Mullerian Inhibitory Substance type II Receptor (MISIIR) gene promoter. Additional transgenic lines have been identified that express the SV40 TAg transgene, but do not develop ovarian tumors. Non-tumor prone mice exhibit typical lifespan for C57Bl/6 mice and are fertile. These mice can be used as syngeneic allograft recipients for tumor cells isolated from Tg-MISIIR-TAg-DR26 mice.
Although tumor imaging is possible, early detection of deep tumors is challenging in small living animals. To enable preclinical studies in an immunologically intact animal model for serous ovarian cancer, we describe a syngeneic mouse model for this type of ovarian cancer that permits in vivo imaging, studies of the tumor microenvironment and tumor immune responses.
We first derived a TAg+ mouse cancer cell line (MOV1) from a spontaneous ovarian tumor harvested in a 26 week-old DR26 Tg-MISIIR-TAg female. Then, we stably transduced MOV1 cells with TurboFP635 Lentivirus mammalian vector that encodes Katushka, a far-red mutant of the red fluorescent protein from sea anemone Entacmaea quadricolor with excitation/emission maxima at 588/635 nm. We orthotopically implanted MOV1(Kat) in the ovary of non-tumor prone Tg-MISIIR-TAg female mice. Tumor progression was followed by in vivo optical imaging and tumor microenvironment was analyzed by immunohistochemistry.
Orthotopically implanted MOV1(Kat) cells developed serous ovarian tumors. MOV1(Kat) tumors could be visualized by in vivo imaging up to three weeks after implantation (fig. 1) and were infiltrated with leukocytes, as observed in human ovarian cancers (fig. 2).
We describe an orthotopic model of ovarian cancer suitable for in vivo imaging of early tumors due to the high pH-stability and photostability of Katushka in deep tissues. We propose the use of this novel syngeneic model of serous ovarian cancer for in vivo imaging studies and monitoring of tumor immune responses and immunotherapies.
Animal Models of Ovarian Cancer Connolly, Denise C.
Cancer Treatment and Research,
01/2009, Letnik:
149
Book Chapter, Journal Article
Recenzirano
As contributors have discussed in previous chapters, our understanding of epithelial ovarian cancer (EOC) biology and clinical management of ovarian cancer patients has improved significantly over ...the years. Yet in spite of all of our advances, the overall improvements in patient outcomes have been incremental. The majority of patients are still diagnosed at advanced stage when the probability of disease recurrence and complications that ultimately result in death are quite high. The typical late-stage diagnosis of EOC is associated with a complex array of genetic and epigenetic alterations present in tumors. As a result, it has been difficult to determine the sequence of events that are involved in tumor initiation, progression, and maintenance. In addition, there are several morphologic variants of EOC including multiple histologic subtypes (e.g., serous, endometrioid, mucinous, clear cell, mixed Muellerian tumors) as well as tumors of low malignant potential.
Purpose: Mullerian inhibiting substance (MIS) is a glycoprotein hormone that causes Mullerian duct regression in male embryos. In
short-term experiments, recombinant human MIS (rhMIS) inhibits ...xenotransplanted human ovarian cancer cell lines that are thought
to be of Mullerian origin. Because this highly lethal cancer has a high recurrence rate after conventional chemotherapy, new
treatments are warranted. We examined whether rhMIS as a novel, nontoxic, naturally occurring growth inhibitor can be an effective
anticancer drug in long-term studies in vivo against allograft tumors that recapitulate human ovarian carcinoma.
Experimental Design: Mouse ovarian carcinoma (MOVCAR) cell lines expressing the early region of the SV40 virus, including the large and small
T-antigen genes under transcriptional control of a portion of the murine MIS receptor type II (MISRII) gene promoter, were
derived from Tg MISIIR-TAg transgenic mice. rhMIS was tested against MOVCAR cells in growth inhibition assays in vitro , and in vivo in 6-week-old female nude mice. Tumor growth in animals was measured at weekly intervals for up to 20 weeks.
Results: MOVCAR cells and tumors express MISRII by Western blot, immunohistochemical, and Northern blot analyses. rhMIS significantly
inhibited MOVCAR cell growth in vitro and in vivo in three separate long-term allotransplantation experiments.
Conclusions: Because rhMIS is an effective anticancer agent in in vitro and in long-term in vivo preclinical experiments against MISRII-positive tumors, we predict that rhMIS can be used safely and effectively to treat
human ovarian malignancies.
Mullerian Inhibiting Substance (MIS), a biological modifier that causes regression of Mullerian ducts in male embryos, is effective as a single agent in vitro and in vivo against human and mouse ...ovarian cancer cell lines expressing MIS type II receptor; however, little is known about how recombinant human MIS (rhMIS), now being scaled for preclinical trials, could be used in combination with cytotoxic or targeted chemotherapeutic agents. Mouse serous and endometrioid ovarian carcinoma cell lines were tested in vitro against rhMIS alone and with doxorubicin, paclitaxel, or cisplatin as agents in clinical use. Because MIS releases FK506 binding protein (FKBP12), which activates the mammalian target of rapamycin (mTOR) downstream of Akt, rhMIS and rapamycin combinations were tested. MIS increases p16 protein levels, and 5'-Aza2'-deoxycytidine (AzadC) induces p16 mRNA; therefore, they were used in combination in vitro and in vivo with a human ovarian cancer cell line. A paclitaxel-resistant human ovarian cancer cell line and its parental line both respond to rhMIS in vitro. Additivity, synergy, or competition was observed with MIS and rapamycin, AzadC, doxorubicin, cisplatin, and paclitaxel, suggesting that MIS in combination with selective targeted therapies might achieve greater activity against ovarian cancer than the use of each individual agent alone. These assays and statistical analyses could be useful in selecting rhMIS and chemotherapeutic agent combinations that enhance clinical efficacy and reduce toxicity.
Aurora kinase A (AURKA) localizes to centrosomes and mitotic spindles where it mediates mitotic progression and chromosomal stability. Overexpression of AURKA is common in cancer, resulting in ...acquisition of alternate non-mitotic functions. In the current study, we identified a novel role for AURKA in regulating ovarian cancer cell dissemination and evaluated the efficacy of an AURKA-selective small molecule inhibitor, alisertib (MLN8237), as a single agent and combined with paclitaxel using an orthotopic xenograft model of epithelial ovarian cancer (EOC). Ovarian carcinoma cell lines were used to evaluate the effects of AURKA inhibition and overexpression on migration and adhesion. Pharmacological or RNA interference-mediated inhibition of AURKA significantly reduced ovarian carcinoma cell migration and adhesion and the activation-associated phosphorylation of the cytoskeletal regulatory protein SRC at tyrosine 416 (pSRC(Y416)). Conversely, enforced expression of AURKA resulted in increased migration, adhesion and activation of SRC in cultured cells. In vivo tumor growth and dissemination were inhibited by alisertib treatment as a single agent. Moreover, combination of alisertib with paclitaxel, an agent commonly used in treatment of EOC, resulted in more potent inhibition of tumor growth and dissemination compared with either drug alone. Taken together, these findings support a role for AURKA in EOC dissemination by regulating migration and adhesion. They also point to the potential utility of combining AURKA inhibitors with taxanes as a therapeutic strategy for the treatment of EOC patients.
Abstract
From a clinical standpoint, epithelial ovarian cancer (EOC) metastasis is a significant cause of morbidity and mortality in patients. The signal transducer and activator of transcription-3 ...(STAT3) protein is a cytoplasmic transcription factor that mediates signals from cytokines and growth factors and non-growth factor tyrosine kinases. Constitutive activation of STAT3 is commonly observed in human ovarian cancer cell lines and primary tumors. In addition to well-established roles for STAT3 in cell proliferation and survival, evidence suggests it also plays a role in tumor cell migration and invasion. To determine the role of STAT3 in ovarian carcinoma cell migration and invasion, we used human ovarian carcinoma cell lines with enforced expression or RNA interference (RNAi)-mediated knockdown of STAT3. To determine the mechanism(s) of STAT3 activation in EOC cells, pharmacologic and RNAi approaches were used to interrogate candidate activators. Specifically, we show that expression of constitutively activated STAT3 results in increased MMP-2 expression and invasion of EOC cells. As Src is both a key mediator of tumor cell migration and invasion and an upstream activator of STAT3, we utilized pharmacologic and RNA interference (RNAi) approaches to determine if Src-mediated activation of STAT3 is essential for migration and invasion in ovarian carcinoma cells. Inhibition of either Src or STAT3 alone similarly diminishes migration of ovarian cancer cells. However, surprisingly, Src inhibition results in a concomitant increase in phosphorylation (activation) of STAT3, suggesting a compensatory mechanism for STAT3 activation. In support of this, simultaneous inhibition of STAT3 and Src results in significantly greater inhibition of migration. To uncover the alternative mechanism for STAT3 activation, we investigated the interactions of STAT3 with key proteins localized in focal adhesion complexes and known to mediate cell motility. This analysis showed that activated STAT3 co-localizes at focal adhesion complexes with phosphorylated focal adhesion kinase (FAK) and neural precursor cell expressed, developmentally downregulated 9 (NEDD9). In addition, using pharmacologic and RNAi-mediated approaches for candidate inhibition, we showed that focal complex localization of STAT3 is dependent on both FAK and NEDD9 and that transcriptional activation of STAT3 in ovarian carcinoma cells is dependent on FAK, rather than JAK2 or SRC. Taken together, these data suggest an important and previously unappreciated role for FAK in STAT3 activation, migration and invasion of ovarian carcinoma cells. These results also nominate FAK as a potential therapeutic target for ovarian cancer treatment.
Citation Format: Fang Xiao, Denise C. Connolly. FAK mediates STAT3 activation, migration and invasion in ovarian carcinoma cells. abstract. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2095. doi:10.1158/1538-7445.AM2014-2095
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