Inhibition of ghrelin action<i>in vitro</i>and<i>in vivo</i>by an RNA-Spiegelmer

Helmling, Steffen; Maasch, Christian; Eulberg, Dirk; Schröder, Werner; Lange, Christian; Vonhoff, Stefan; Wlotzka, Britta; Tschöp, Matthias H.; Rosewicz, Stefan; Klussmann, Sven

doi:10.1073/pnas.0404175101

Cited by 137 publications

(100 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Both ghrelin and GHS-R are being investigated as therapeutic targets (Schellekens et al, 2010). In vitro generated biostable RNA-based compounds that specifically bind ghrelin have been used in animal models to inhibit ghrelin-mediated GHS-R activation (Helmling et al, 2004). Zorilla et al have developed an anti-ghrelin vaccine, providing a novel method to block ghrelin action (Zorrilla et al, 2006).…”

Section: Ghrelin and Obesitymentioning

confidence: 99%

Ghrelin in obesity and endocrine diseases

Scerif

Goldstone

Korbonits

2011

Molecular and Cellular Endocrinology

View full text Add to dashboard Cite

Section: Ghrelin and Obesitymentioning

confidence: 99%

Ghrelin in obesity and endocrine diseases

Scerif

Goldstone

Korbonits

2011

Molecular and Cellular Endocrinology

View full text Add to dashboard Cite

“…11 Hence, Spiegelmers show excellent biostability without any further chemical modifications, which renders then very well suited for in vitro and in vivo applications. [12][13][14][15] We therefore hypothesized that Spiegelmer-based blockade of CCL2 would be suitable for the treatment of lupus nephritis and other disease manifestations of systemic lupus erythematosus. Here we report the identification of the Spiegelmer mNOX-E36 that specifically inhibits murine CCL2 (mCCL2) in vitro in the absence of type I IFN induction in dendritic cells, recently described for natural and synthetic RNA.…”

mentioning

confidence: 99%

Spiegelmer Inhibition of CCL2/MCP-1 Ameliorates Lupus Nephritis in MRL-(Fas)lpr Mice

Kulkarni

Pawar

Purschke

et al. 2007

Journal of the American Society of Nephrology

Self Cite

168

124

View full text Add to dashboard Cite

The monocyte chemoattractant protein CCL2 is crucial for monocyte and T cell recruitment from the vascular to the extravascular compartment at sites of inflammation. CCL2 is expressed in human lupus nephritis and was shown to mediate experimental lupus; therefore, CCL2 antagonists may be beneficial for therapy. This study describes the L-enantiomeric RNA oligonucleotide mNOX-E36, a so-called Spiegelmer that binds murine CCL2 with high affinity and neutralizes its action in vitro and in vivo. The mirror image configuration of the Spiegelmer confers nuclease resistance and thus excellent biostability. mNOX-E36 does not induce type I IFN via Toll-like receptor-7 or cytosolic RNA receptors, as recently shown for certain synthetic D-RNA. Autoimmune-prone MRL lpr/lpr mice that were treated with a polyethylene glycol form of mNOX-E36 from weeks 14 to 24 of age showed prolonged survival associated with a robust improvement of lupus nephritis, peribronchial inflammation, and lupus-like inflammatory skin lesions. Thus, mNOX-E36 -based inhibition of CCL2 represents a novel strategy for the treatment of autoimmune tissue injury, such as lupus nephritis.

show abstract

“…In Vitro Selection and Pulldown Assays-Automated and manual in vitro selection experiments were performed against the biotinylated 21-amino acid mirror-image peptide, representing a central sequence of HMGA1a/b in selection buffer resembling intracellular ion conditions and pH (25 mM TrisHCl, pH 7.0, 140 mM KCl, 12 mM NaCl, 0.8 mM MgCl 2 , and 0.1% Tween 20) at 37°C (13,23,24). Pulldown assays were carried out as described (13,23,24).…”

Section: Methodsmentioning

confidence: 99%

“…Spiegelmers (German: Spiegel ϭ mirror) are structured biostable L-oligonucleotides that differ from their aptamer counterparts in their sugar moiety, which consists of mirrorimage L-(deoxy)ribose rather than D-(deoxy)ribose and makes Spiegelmers highly resistant to nucleases (11,12). Spiegelmers have already been described to act potently as inhibitors in vivo (13)(14)(15) and have proven to be exceptionally safe in two Phase I clinical studies. 4 Here, we report the generation of a Spiegelmer, binding to HMGA1 with high affinity.…”

Section: From Noxxon Pharma Ag Max-dohrn-strasse 8-10 D-10589 Berlimentioning

confidence: 99%

Polyetheylenimine-Polyplexes of Spiegelmer NOX-A50 Directed against Intracellular High Mobility Group Protein A1 (HMGA1) Reduce Tumor Growth in Vivo

Maasch¹,

Vater²,

Purschke

et al. 2010

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

High mobility group A1 (HMGA1) proteins belong to a group of architectural transcription factors that are overexpressed in a range of human malignancies, including pancreatic adenocarcinoma. They promote anchorage-independent growth and epithelial-mesenchymal transition and are therefore suggested as potential therapeutic targets. Employing in vitro selection techniques against a chosen fragment of HMGA1, we have generated biostable L-RNA oligonucleotides, so-called Spiegelmers, that specifically bind HMGA1b with low nanomolar affinity. We demonstrate that the best binding Spiegelmers, NOX-A50 and NOX-f33, compete HMGA1b from binding to its natural binding partner, AT-rich doublestranded DNA. We describe a formulation method based on polyplex formation with branched polyethylenimine for efficient delivery of polyethylene glycol-modified Spiegelmers and show improved tissue distribution and persistence in mice. In a xenograft mouse study using the pancreatic cancer cell line PSN-1, subcutaneous administration of 2 mg/kg per day NOX-A50 formulated in polyplexes showed an enhanced delivery of NOX-A50 to the tumor and a significant reduction of tumor volume. Our results demonstrate that intracellular targets can be successfully addressed with a Spiegelmer using polyethylenimine-based delivery and underline the importance of HMGA1 as a therapeutic target in pancreatic cancer.The mammalian nonhistone chromatin high mobility group A1 proteins HMGA1a, 3 HMGA1b, and HMGA1c belong to one protein family encoded by the HMGA1 gene. They act as architectural transcription factors by binding via AT hook motifs to the minor groove of AT-rich DNA, leading to modulation of chromatin and nucleosome remodeling. Their ability to regulate the activity of several genes through the recognition and alteration of the DNA double helix and chromatin substrates has been linked to the development of human cancers (1-3). The overexpression of HMGA proteins is associated with different types of tumors, e.g. pancreas, breast, prostate, cervical, gastric, colorectal cancer as well as lymphomas and neuroblastic malignancies (4 -6). The level of expression of HMGA1a/b is low or almost undetectable in most differentiated or nonproliferating normal cells (7-9) but is rapidly up-regulated in response to growth-stimulatory factors (8, 10). Thus, HMGA1 is considered a promising target to treat cancer development, progression, and metastasis.The aim of our study was to evaluate whether the intracellular HMGA1 protein family can be successfully targeted with a Spiegelmer, a large structured molecule that is comparable with a monoclonal antibody in terms of specific target binding. Spiegelmers (German: Spiegel ϭ mirror) are structured biostable L-oligonucleotides that differ from their aptamer counterparts in their sugar moiety, which consists of mirrorimage L-(deoxy)ribose rather than D-(deoxy)ribose and makes Spiegelmers highly resistant to nucleases (11,12). Spiegelmers have already been described to act potently as inhibitors in vivo (13-15) and have ...

show abstract

Inhibition of ghrelin actionin vitroandin vivoby an RNA-Spiegelmer

Cited by 137 publications

References 53 publications

Ghrelin in obesity and endocrine diseases

Ghrelin in obesity and endocrine diseases

Spiegelmer Inhibition of CCL2/MCP-1 Ameliorates Lupus Nephritis in MRL-(Fas)lpr Mice

Polyetheylenimine-Polyplexes of Spiegelmer NOX-A50 Directed against Intracellular High Mobility Group Protein A1 (HMGA1) Reduce Tumor Growth in Vivo

Contact Info

Product

Resources

About