Plasma Membrane Nucleolin Is a Receptor for the Anticancer Aptamer AS1411 in MV4-11 Leukemia Cells

Soundararajan, Sridharan; Wang, Li; Sridharan, Vijayalakshmi; Chen, Weiwei; Courtenay-Luck, Nigel; Jones, David R.; Spicer, Eleanor K.; Fernandes, Daniel

doi:10.1124/mol.109.055947

Cited by 204 publications

(165 citation statements)

References 25 publications

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“…Recent studies showed that nucleolin, a nuclear protein, was present on the surface of various cell types, despite lacking a transmembrane domain or a signal sequence (24,25). Therefore, nucleolin acts as a receptor on plasma membrane and shuttles the ligand from the plasma membrane to the cytoplasm and nucleus (24,26). Likewise, KLF4 may also migrate to the cell surface and act as a kallistatin-binding protein.…”

Section: Discussionmentioning

confidence: 99%

Kruppel-like Factor 4 Is a Novel Mediator of Kallistatin in Inhibiting Endothelial Inflammation via Increased Endothelial Nitric-oxide Synthase Expression

Shen

Smith

Hsu

et al. 2009

Journal of Biological Chemistry

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Kallistatin is a plasma protein that exhibits pleiotropic effects in vasodilation, anti-angiogenesis, and anti-inflammation. To isolate a kallistatin-binding protein that mediates the vascular actions of kallistatin, we screened and identified a positive clone from a human heart cDNA expression library by using an alkaline phosphatase-kallistatin fusion protein binding assay. Sequence analysis revealed that kallistatin-binding protein is human Kruppel-like factor 4 (KLF4). KLF4 was localized on the plasma membrane of HEK-293 cells and endothelial cells overexpressing KLF4. KLF4 and kallistatin complex formation was identified in endothelial cells by immunoprecipitation followed by immunoblotting. We showed that kallistatin inhibits tumor necrosis factor-␣-induced NF-B activation, as well as vascular cell adhesion molecule-1 and monocyte chemoattractant protein-1 expression in endothelial cells, whereas knockdown of KLF4 by small interfering RNA oligonucleotide abolished the effect of kallistatin. Kallistatin increased endothelial nitric-oxide synthase (eNOS) expression and nitric oxide levels, and these effects were also blocked by KLF4 small interfering RNA oligonucleotide. Moreover, inhibition of eNOS by RNA interference or by NOS inhibitor abolished the blocking effect of kallistatin on vascular cell adhesion molecule-1 and monocyte chemoattractant protein-1 expression. In summary, we identified KLF4 as a kallistatin-binding protein, which has a novel role in mediating the anti-inflammatory actions of kallistatin via increasing eNOS expression in endothelial cells. This study provides a new target for modulating endothelial function in vascular disease.Kallistatin was discovered as a tissue kallikrein inhibitor from human plasma (1, 2). Recent studies indicate that kallistatin exerts pleiotropic effects such as vasodilation and inhibition of angiogenesis, tumor growth, inflammation, and oxidative stress, independent of tissue kallikrein (3-7). We showed that an intravenous bolus injection of human kallistatin caused a rapid and transient reduction of mean arterial blood pressure in anesthetized rats and that kallistatin induced relaxation in isolated aorta rings (3). Moreover, a high affinity binding site for kallistatin was identified in the aortic membrane using 125 Ilabeled kallistatin (3). Furthermore, kallistatin directly stimulated the proliferation and migration of cultured vascular smooth muscle cells by activating mitogen-activated protein kinases (8). These findings suggest the presence of kallistatin receptors or binding proteins that mediate its effects in the vasculature. However, the identity of the kallistatin receptor/ binding protein has not been determined.Kallistatin is a negative acute-phase protein, which is reduced after acute lipopolysaccharide-induced inflammation and in chronic inflammatory disease (9). Transgenic mice overexpressing kallistatin are more resistant to lipopolysaccharideinduced lethality (10). Kallistatin administration via gene delivery significantly decreased neutroph...

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Section: Discussionmentioning

confidence: 99%

Kruppel-like Factor 4 Is a Novel Mediator of Kallistatin in Inhibiting Endothelial Inflammation via Increased Endothelial Nitric-oxide Synthase Expression

Shen

Smith

Hsu

et al. 2009

Journal of Biological Chemistry

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“…The widely demonstrated role of NCL in tumorigenesis suggests that inhibition of its oncogenic actions reduces tumor aggressiveness (9,13,19,24,32,34,47), and several studies have proposed NCL as an ideal target for antineoplastic therapies in different solid and hematological malignancies (9,19,25,34,37,38,41,42,48,49). Given the selective presence of NCL on cancer cells and cancer-associated endothelial cells, but not on normal cells, molecules targeting NCL might represent an effective approach for the selective delivery of drugs or toxins to tumors while minimizing side effects (25,41).…”

Section: Discussionmentioning

confidence: 99%

“…NCL is able to shuttle between the cell surface and the cytoplasm of cancer cells (32,34), a property that makes NCL an attractive target for the selective delivery of antineoplastic drugs or prodrugs, leaving normal cells unaffected (25).…”

Section: Significancementioning

confidence: 99%

Human anti-nucleolin recombinant immunoagent for cancer therapy

Palmieri

Richmond

Piovan

et al. 2015

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

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Nucleolin (NCL) is a nucleocytoplasmic protein involved in many biological processes, such as ribosomal assembly, rRNA processing, and mRNA stabilization. NCL also regulates the biogenesis of specific microRNAs (miRNAs) involved in tumor development and aggressiveness. Interestingly, NCL is expressed on the surface of actively proliferating cancer cells, but not on their normal counterparts. Therefore, NCL is an attractive target for antineoplastic treatments. Taking advantage of phage-display technology, we engineered a fully human single-chain fragment variable, named 4LB5. This immunoagent binds NCL on the cell surface, it is translocated into the cytoplasm of target cells, and it abrogates the biogenesis of NCLdependent miRNAs. Binding of 4LB5 to NCL on the cell surface of a variety of breast cancer and hepatocellular carcinoma cell lines, but not to normal-like MCF-10a breast cells, dramatically reduces cancer cell viability and proliferation. Finally, in orthotopic breast cancer mouse models, 4LB5 administration results in a significant reduction of the tumor volume without evident side effects. In summary, here we describe, to our knowledge, the first anti-NCL single-chain fragment variable displaying antineoplastic activity against established solid tumors, which could represent the prototype of novel immunebased NCL-targeting drugs with clinical potential as diagnostic and therapeutic tools in a wide variety of human cancers.is one of the most abundant nonribosomal proteins in the nucleolus (1), first identified in ribosomal RNA processing (2). Additional studies have demonstrated that NCL is a multifunctional nucleocytoplasmic protein, involved in ribosomal assembly, chromatin decondensation, transcription, nucleo-cytoplasmic import/export, and chromatin remodeling (3, 4). NCL is frequently up-regulated in cancer and in cancerassociated endothelial cells compared with normal tissues (5, 6), where it is also present on the cell surface (7,8). Altered NCL expression and localization results in oncogenic effects, such as stabilization of AKT, Bcl-2, Bcl-XL, and IL-2 mRNAs (9-11). Moreover, surface-NCL acts as a receptor for several oncogenic ligands (12-15) and viruses (16). Recently, we reported that NCL has a critical protumorigenic function regulating the biogenesis of selected microRNAs (miRNAs), a class of noncoding single-stranded RNAs 19-22 nt in length (17) that regulate gene expression at the posttranscriptional level by targeting mRNAs in a sequence-specific manner (18). In fact, NCL enhances the maturation of specific miRNAs (including miR-21, miR-221, and miR-222) causally involved in cancer pathogenesis and resistance to several antineoplastic treatments (19-23). Our findings demonstrated that NCL modulates the biogenesis of these miRNAs at the posttranscriptional level, enhancing their maturation from pri-to premiRNAs, identifying a novel NCL-dependent oncogenic mechanism (19).Because of its oncogenic role and specific expression on cancer cells surface, NCL represents an attractive target f...

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“…It controls the metabolism of DNA and RNA in the nucleolus (44,45), shuttles proteins into the nucleus (46), provides posttranscriptional regulation of strategic mRNAs in the cytoplasm (47,48), and serves as a ligand for proteins, such as midkine and the heparin-binding growth-associated molecule, on the cell surface (49). NCL is also involved in the proliferation of many types of cancer (34,(50)(51)(52) and serves as a prognosis marker and therapeutic target for cancer treatment (53)(54)(55). Furthermore, NCL also participates in the pathogenic mechanism and mediates the replication of several viruses and/or serves as a cell surface receptor for Escherichia coli O157 (56), Helicobacter pylori (57), RSV (28), adeno-associated virus type-2 (AAV-2) (58), coxsackie B virus (59), and HIV (29).…”

Section: Discussionmentioning

confidence: 99%

Cell Surface Nucleolin Facilitates Enterovirus 71 Binding and Infection

Wang

Huang

et al. 2015

J Virol

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Because the pathogenesis of enterovirus 71 (EV71) remains mostly ambiguous, identifying the factors that mediate viral binding and entry to host cells is indispensable to ultimately uncover the mechanisms that underlie virus infection and pathogenesis. Despite the identification of several receptors/attachment molecules for EV71, the binding, entry, and infection mechanisms of EV71 remain unclear. Herein, we employed glycoproteomic approaches to identify human nucleolin as a novel binding receptor for EV71. Glycoproteins purified by lectin chromatography from the membrane extraction of human cells were treated with sialidase, followed by immunoprecipitation with EV71 particles. Among the 16 proteins identified by tandem mass spectrometry analysis, cell surface nucleolin attracted our attention. We found that EV71 interacted directly with nucleolin via the VP1 capsid protein and that an antinucleolin antibody reduced the binding of EV71 to human cells. In addition, the knockdown of cell surface nucleolin decreased EV71 binding, infection, and production in human cells. Furthermore, the expression of human nucleolin on the cell surface of a mouse cell line increased EV71 binding and conferred EV71 infection and production in the cells. These results strongly indicate that human nucleolin can mediate EV71 binding to and infection of cells. Our findings also demonstrate that the use of glycoproteomic approaches is a reliable methodology to discover novel receptors for pathogens. IMPORTANCEOutbreaks of EV71 have been reported in Asia-Pacific countries and have caused thousands of deaths in young children during the last 2 decades. The discovery of new EV71-interacting molecules to understand the infection mechanism has become an emergent issue. Hence, this study uses glycoproteomic approaches to comprehensively investigate the EV71-interacting glycoproteins. Several EV71-interacting glycoproteins are identified, and the role of cell surface nucleolin in mediating the attachment and entry of EV71 is characterized and validated. Our findings not only indicate a novel target for uncovering the EV71 infection mechanism and anti-EV71 drug discovery but also provide a new strategy for virus receptor identification.

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Plasma Membrane Nucleolin Is a Receptor for the Anticancer Aptamer AS1411 in MV4-11 Leukemia Cells

Cited by 204 publications

References 25 publications

Kruppel-like Factor 4 Is a Novel Mediator of Kallistatin in Inhibiting Endothelial Inflammation via Increased Endothelial Nitric-oxide Synthase Expression

Kruppel-like Factor 4 Is a Novel Mediator of Kallistatin in Inhibiting Endothelial Inflammation via Increased Endothelial Nitric-oxide Synthase Expression

Human anti-nucleolin recombinant immunoagent for cancer therapy

Cell Surface Nucleolin Facilitates Enterovirus 71 Binding and Infection

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