Pyridopyrimidine derivatives as inhibitors of cyclic nucleotide synthesis: Application for treatment of diarrhea

Kots, Alexander Y.; Choi, Byron; Estrella-Jimenez, Maria E.; Warren, Cirle A.; Gilbertson, Scott R.; Guerrant, Richard L.; Murad, Ferid

doi:10.1073/pnas.0803096105

Cited by 63 publications

(46 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Oct3=4 and NANOG proteins are markers of pluripotency [24], and their staining was drastically reduced in differentiated cells. On the contrary, expression of a1 and to the validation based on established cell models: sGCpositive BE2 human neuroblastoma cells [17], and sGCnegative T84 human colorectal carcinoma cells [25] and COS7 [25] (see Supplementary Fig. S1; Supplementary Data are available online at www.liebertonline.com=scd).…”

Section: Resultsmentioning

confidence: 99%

Nitric Oxide Receptor Soluble Guanylyl Cyclase Undergoes Splicing Regulation in Differentiating Human Embryonic Cells

Sharin

Mujoo

Kots

et al. 2011

Stem Cells and Development

Self Cite

View full text Add to dashboard Cite

Nitric oxide (NO), an important mediator molecule in mammalian physiology, initiates a number of signaling mechanisms by activating the enzyme soluble guanylyl cyclase (sGC). Recently, a new role for NO=cyclic guanosine monophosphate signaling in embryonic development and cell differentiation has emerged. The changes in expression of NO synthase isoforms and various sGC subunits has been demonstrated during human and mouse embryonic stem (ES) cells differentiation. Previously, our laboratory demonstrated that nascent a1 sGC transcript undergoes alternative splicing and that expression of a1 sGC splice forms directly affects sGC activity. Expression of sGC splice variants in the process of human ES (hES) cells differentiation has not been investigated. In this report, we demonstrate that a1 sGC undergoes alternative splicing during random hES differentiation for the first time. Our results indicate that C-a1 sGC splice form is expressed at high levels in differentiating cells and its intracellular distribution varies from canonical a1 sGC subunit. Together, our data suggest that alternative splicing of sGC subunits is associated with differentiation of hES cells.

show abstract

Section: Resultsmentioning

confidence: 99%

Nitric Oxide Receptor Soluble Guanylyl Cyclase Undergoes Splicing Regulation in Differentiating Human Embryonic Cells

Sharin

Mujoo

Kots

et al. 2011

Stem Cells and Development

Self Cite

View full text Add to dashboard Cite

show abstract

“…Then, an NO donor, spermine NONOate (0.1 mM) or forskolin (10 M) was added and incubation continued for another 10 min. Medium was aspirated and cGMP/cAMP was extracted by rapid freezing of the plates at Ϫ80°C in the presence of 50 mM sodium acetate, pH 4.0, (0.3 ml per well), then measured by enzyme-linked immunosorbent assay as described previously (Kots et al, 2008). For the assay of the basal level, tumor cells were harvested without IBMX preincubation and further treatments.…”

Section: Methodsmentioning

confidence: 99%

Restoring Soluble Guanylyl Cyclase Expression and Function Blocks the Aggressive Course of Glioma

Zhu¹,

Li²,

Zheng³

et al. 2011

Mol Pharmacol

Self Cite

View full text Add to dashboard Cite

The NO and cGMP signaling pathways are of broad physiological and pathological significance. We compared the NO/soluble guanylyl cyclase (sGC)/cGMP pathway in human glioma tissues and cell lines with that of healthy control samples and demonstrated that sGC expression is significantly lower in glioma preparations. Our analysis of GEO databases (National Cancer Institute) further revealed a statistically significant reduction of sGC transcript levels in human glioma specimens. On the other hand, the expression levels of particulate (membrane) guanylyl cyclases (pGC) and cGMP-specific phosphodiesterase (PDE) were intact in the glioma cells that we have tested. Pharmacologically manipulating endogenous cGMP generation in glioma cells through either stimulating pGC by ANP/BNP, or blocking PDE by 3-isobutyl-1-methylxanthine/ zaprinast caused significant inhibition of proliferation and colony formation of glioma cells. Genetically restoring sGC expression also correlated inversely with glioma cells growth. Orthotopic implantation of glioma cells transfected with an active mutant form of sGC (sGC␣1␤1 Cys105 ) in athymic mice increased the survival time by 4-fold over the control. Histological analysis of xenografts overexpressing ␣1␤1Cys105 sGC revealed changes in cellular architecture that resemble the morphology of normal cells. In addition, a decrease in angiogenesis contributed to glioma inhibition by sGC/cGMP therapy. Our study proposes the new concept that suppressed expression of sGC, a key enzyme in the NO/cGMP pathway, may be associated with an aggressive course of glioma. The sGC/ cGMP signaling-targeted therapy may be a favorable alternative to chemotherapy and radiotherapy for glioma and perhaps other tumors.

show abstract

“…cGMP was extracted on ice for 30 min. Supernatant fractions were neutralized with potassium carbonate (12.5 L of 2 M) and used for cGMP assay by ELISA as described (28). Pellets were dissolved in 0.5 mL of 0.1 M NaOH and used to measure protein by the BioRad dye-binding assay.…”

Section: Treatment Of Partially Differentiated Cells With the Activatmentioning

confidence: 99%

Role of nitric oxide signaling components in differentiation of embryonic stem cells into myocardial cells

Mujoo

Sharin

Bryan

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

View full text Add to dashboard Cite

cardiomyogenesis ͉ cyclic GMP ͉ soluble guanylyl cyclase ͉ NO donors ͉ sGC activators E mbryonic stem (ES) cells are derived from the inner cell mass of the preimplantation embryo, and because of their selfrenewal and pluripotency, they are thought to revolutionize the field of regenerative medicine (1-3). Although transplantation of stem cells into animal models of cardiac injury and Parkinson's disease has revealed some beneficial effects, a better understanding of the role of specific signaling pathways involved in proliferation and differentiation of ES cells is necessary to improve their use in clinical medicine (4-6). Components of a number of signaling pathways such as Wnt/␤-catenin (7), phosphotidyle-inosital 3-kinase (8, 9), MAPK (10), and NO (11) have been shown to regulate proliferation and differentiation of stem cells.NO is a diffusible short-lived free radical and a signaling molecule with a number of important physiological functions such as smooth muscle relaxation, neurotransmission, and inhibition of platelet aggregation and host defense mechanisms (12). It also plays a role in the pathology of several inflammatory diseases and other pathological conditions such as cancer, diabetes, and neurodegenerative diseases (13,14). NO plays an important role in the control of heart rate, contractibility, coronary perfusion, and cardiac development (15, 16). It is synthesized by enzymes called nitric oxide synthases such as NOS-1 (neuronal NOS), NOS-2 (inducible NOS), and NOS-3 (endothelial NOS) that catalyze the oxidation of L-arginine into L-citruline with the release of NO. NO can also be measured in biological systems as metabolites of NO and as nitrites and nitrates (17). The NO receptor soluble guanylyl cyclase (sGC) is a heme-containing heterodimer with ␣ and ␤ subunits (12). Binding of NO to the heme prosthetic group of sGC catalyzes the conversion of GTP into the second messenger cGMP that can exert many physiological effects, such as mediating vascular smooth muscle tone and motility, phototransduction, and maintenance of fluid and electrolyte homeostasis by interaction of cGMP with downstream effectors such as a family of cGMP-dependent protein kinases, cGMP-dependent phosphodiesterases, and cyclic nucleotide gated channels (11).Our previous studies with mouse and human ES cells demonstrated a time-dependent increase in mRNA and protein levels of different subunits of sGC (with the exception of ␤2 mRNA in H-9 cells) during both mouse and human ES cell differentiation into cells of cardiac lineage. Compared with NOS-1, NOS-2, and NOS-3 mRNA and protein levels were also induced during ES cell differentiation (18,19), thereby demonstrating the involvement of NO signaling components during differentiation of ES cells into cardiac cells. Previous studies by other investigators (20) have shown increased cardiomyogenesis with NO donors and by delivery of the NOS-2 gene in mouse ES cells.In this study, we demonstrate the role of NO and cGMP in differentiation of human and mouse ES cells by regulating t...

show abstract

Pyridopyrimidine derivatives as inhibitors of cyclic nucleotide synthesis: Application for treatment of diarrhea

Abstract: adenylyl cyclase ͉ cGMP ͉ guanylyl cyclase ͉ cAMP ͉ toxin-induced diarrhea

Cited by 63 publications

References 27 publications

Nitric Oxide Receptor Soluble Guanylyl Cyclase Undergoes Splicing Regulation in Differentiating Human Embryonic Cells

Nitric Oxide Receptor Soluble Guanylyl Cyclase Undergoes Splicing Regulation in Differentiating Human Embryonic Cells

Restoring Soluble Guanylyl Cyclase Expression and Function Blocks the Aggressive Course of Glioma

Role of nitric oxide signaling components in differentiation of embryonic stem cells into myocardial cells

Contact Info

Product

Resources

About