A Mixed Mirror-image DNA/RNA Aptamer Inhibits Glucagon and Acutely Improves Glucose Tolerance in Models of Type 1 and Type 2 Diabetes

Vater, Axel; Sell, Simone; Kaczmarek, Przemysław; Maasch, Christian; Buchner, Klaus; Pruszyńska‐Oszmałek, Ewa; Kołodziejski, Paweł; Purschke, Werner G.; Nowak, Krzysztof W.; Strowski, Mathias Z.; Klussmann, Sven

doi:10.1074/jbc.m112.444414

Cited by 53 publications

(38 citation statements)

References 39 publications

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“…Similarly, glucagon receptor antibodies have been shown to lower hyperglycemia in T2D (15), but this is the first demonstration, to our knowledge, of normalization of glycemia and HbA1c in complete insulin deficiency, a result that, to our knowledge, has never been achieved with existing therapies. Previous neutralization of glucagon itself reduced, but did not normalize, blood glucose levels (27)(28)(29). Synthetic glucagon receptor antagonists (30-33) similarly improved the diabetes but did not normalize it.…”

Section: Discussionmentioning

confidence: 89%

Glucagon receptor antibody completely suppresses type 1 diabetes phenotype without insulin by disrupting a novel diabetogenic pathway

Wang

Yan²,

Shi³

et al. 2015

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

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Insulin monotherapy can neither maintain normoglycemia in type 1 diabetes (T1D) nor prevent the long-term damage indicated by elevated glycation products in blood, such as glycated hemoglobin (HbA1c). Here we find that hyperglycemia, when unaccompanied by an acute increase in insulin, enhances itself by paradoxically stimulating hyperglucagonemia. Raising glucose from 5 to 25 mM without insulin enhanced glucagon secretion ∼two-to fivefold in InR1-G9 α cells and ∼18-fold in perfused pancreata from insulindeficient rats with T1D. Mice with T1D receiving insulin treatment paradoxically exhibited threefold higher plasma glucagon during hyperglycemic surges than during normoglycemic intervals. Blockade of glucagon action with mAb Ac, a glucagon receptor (GCGR) antagonizing antibody, maintained glucose below 100 mg/dL and HbA1c levels below 4% in insulin-deficient mice with T1D. In rodents with T1D, hyperglycemia stimulates glucagon secretion, up-regulating phosphoenolpyruvate carboxykinase and enhancing hyperglycemia. GCGR antagonism in mice with T1D normalizes glucose and HbA1c, even without insulin.glucagon receptor | antibody | type 1 diabetes | insulin

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Section: Discussionmentioning

confidence: 89%

Glucagon receptor antibody completely suppresses type 1 diabetes phenotype without insulin by disrupting a novel diabetogenic pathway

Wang

Yan²,

Shi³

et al. 2015

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

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“…In the case of antibody drug conjugates, which utilize this linkage, the long systemic circulation half-lives of these molecules, *10-20 days [27,28], has led investigators to utilize hindered disulfides that bear a methyl group adjacent to one or both sides of the disulfide linkage to further increase serum stability [26,27]. However, aptamers display clearance rates in minutes [29] which can be extended by conjugation to high molecular weight (20-40 kDa) PEG, which have reported half-lives ranging from 3 to 12 h [29][30][31]. rGel has a molecular weight of 28 kDa, thus we would expect our aptamerrGel conjugates to behave similarly.…”

Section: Discussionmentioning

confidence: 99%

Improved Synthesis andIn VitroEvaluation of an Aptamer Ribosomal Toxin Conjugate

Kelly

Kratschmer

Maier

et al. 2016

Nucleic Acid Therapeutics

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Delivery of toxins, such as the ricin A chain, Pseudomonas exotoxin, and gelonin, using antibodies has had some success in inducing specific toxicity in cancer treatments. However, these antibody-toxin conjugates, called immunotoxins, can be bulky, difficult to express, and may induce an immune response upon in vivo administration. We previously reported delivery of a recombinant variant of gelonin (rGel) by the full-length prostate-specific membrane antigen (PSMA) binding aptamer, A9, to potentially circumvent some of these problems. Here, we report a streamlined approach to generating aptamer-rGel conjugates utilizing a chemically synthesized minimized form of the A9 aptamer. Unlike the full-length A9 aptamer, this minimized variant can be chemically synthesized with a 5¢ terminal thiol. This facilitates the large scale synthesis and generation of aptamer toxin conjugates linked by a reducible disulfide linkage. Using this approach, we generated aptamertoxin conjugates and evaluated their binding specificity and toxicity. On PSMA(+) LNCaP prostate cancer cells, the A9.min-rGel conjugate demonstrated an IC 50 of *60 nM. Additionally, we performed a stability analysis of this conjugate in mouse serum where the conjugate displayed a t 1/2 of *4 h, paving the way for future in vivo experiments.

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“…The isolated aptamer sequences can be chemically synthesised as L-RNA stereoisomers (''Spigelmers''), that are resistant to nuclease cleavage and bind specifically to the natural target (composed of L-amino acids) [9]. Vater et al and Purschke et al have demonstrated that mixed L-DNA/L-RNA Spiegelmers have sufficient stability to mediate clinically-relevant biological effects in vivo, the specific inhibition of glucagon in mouse models of type 1 and type 2 diabetes [10], and neutralisation of sphingosine-1-phosphate, a mediator of angiogenesis [11]. In future it may be possible to evolve Spiegelmers directly using polymerases synthesised from D-amino acids.…”

Section: Stabilitymentioning

confidence: 98%

Towards applications of synthetic genetic polymers in diagnosis and therapy

Taylor

Arangundy-Franklin

Holliger

2014

Current Opinion in Chemical Biology

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A Mixed Mirror-image DNA/RNA Aptamer Inhibits Glucagon and Acutely Improves Glucose Tolerance in Models of Type 1 and Type 2 Diabetes

Cited by 53 publications

References 39 publications

Glucagon receptor antibody completely suppresses type 1 diabetes phenotype without insulin by disrupting a novel diabetogenic pathway

Glucagon receptor antibody completely suppresses type 1 diabetes phenotype without insulin by disrupting a novel diabetogenic pathway

Improved Synthesis andIn VitroEvaluation of an Aptamer Ribosomal Toxin Conjugate

Towards applications of synthetic genetic polymers in diagnosis and therapy

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