Top

Diabetologia

Published in:

Open Access 01-07-2018 | Article

Evaluation of anti-insulin receptor antibodies as potential novel therapies for human insulin receptoropathy using cell culture models

Authors: Gemma V. Brierley, Kenneth Siddle, Robert K. Semple

Published in: Diabetologia | Issue 7/2018

Abstract

Aims/hypothesis

Bi-allelic loss-of-function mutations in the INSR gene (encoding the insulin receptor [INSR]) commonly cause extreme insulin resistance and early mortality. Therapeutic options are limited, but anti-INSR antibodies have been shown to activate two mutant receptors, S323L and F382V. This study evaluates four well-characterised murine anti-INSR monoclonal antibodies recognising distinct epitopes (83-7, 83-14, 18-44, 18-146) as surrogate agonists for potential targeted treatment of severe insulin resistance arising from insulin receptoropathies.

Methods

Ten naturally occurring mutant human INSRs with defects affecting different aspects of receptor function were modelled and assessed for response to insulin and anti-INSR antibodies. A novel 3T3-L1 adipocyte model of insulin receptoropathy was generated, permitting conditional knockdown of endogenous mouse Insr by lentiviral expression of species-specific short hairpin (sh)RNAs with simultaneous expression of human mutant INSR transgenes.

Results

All expressed mutant INSR bound to all antibodies tested. Eight mutants showed antibody-induced autophosphorylation, while co-treatment with antibody and insulin increased maximal phosphorylation compared with insulin alone. After knockdown of mouse Insr and expression of mutant INSR in 3T3-L1 adipocytes, two antibodies (83-7 and 83-14) activated signalling via protein kinase B (Akt) preferentially over signalling via extracellular signal-regulated kinase 1/2 (ERK1/2) for seven mutants. These antibodies stimulated glucose uptake via P193L, S323L, F382V and D707A mutant INSRs, with antibody response greater than insulin response for D707A.

Conclusions/interpretation

Anti-INSR monoclonal antibodies can activate selected naturally occurring mutant human insulin receptors, bringing closer the prospect of novel therapy for severe insulin resistance caused by recessive mutations.

Available only for authorised users

Ebina Y, Ellis L, Jarnagin K et al (1985) The human insulin receptor cDNA: the structural basis for hormone-activated transmembrane signalling. Cell 40:747–758CrossRefPubMed

Ullrich A, Bell R, Chen EY et al (1985) Human insulin receptor and its relationship to the tyrosine kinase family of oncogenes. Nature 313:756–761CrossRefPubMed

Yoshimasa Y, Seino S, Whittaker J et al (1988) Insulin-resistant diabetes due to a point mutation that prevents insulin proreceptor processing. Science 240:784–787CrossRefPubMed

Kadowaki T, Bevins CL, Cama A et al (1988) Two mutant alleles of the insulin receptor gene in a patient with extreme insulin resistance. Science 240:787–790CrossRefPubMed

Semple RK, Savage DB, Brierley GV, O’Rahilly S (2016) Syndromes of severe insulin resistance and/or lipodystrophy. In: Weiss RE, Refetoff S (eds) Genetic diagnosis of endocrine disorders, 2nd edn. Academic Press, Cambridge, MA, pp 307–324CrossRef

Taylor SI, Cama A, Accili D et al (1992) Mutations in the insulin receptor gene. Endocr Rev 13:566–595CrossRefPubMed

Lebrun C, Baron V, Kaliman P et al (1993) Antibodies to the extracellular receptor domain restore the hormone-insensitive kinase and conformation of the mutant insulin receptor valine 382. J Biol Chem 268:11272–11277PubMed

Krook A, Soos M, Kumar S, Siddle K, O’Rahilly S (1996) Functional activation of mutant human insulin receptor by monoclonal antibody. Lancet 347:1586–1590CrossRefPubMed

Ayyar BV, Arora S, O’Kennedy R (2016) Coming-of-age of antibodies in cancer therapeutics. Trends Pharmacol Sci 37:1009–1028CrossRefPubMed

10.

Reichert JM (2016) Antibodies to watch in 2017. MAbs 1–61

11.

Johnson KW, Neale A, Gordon A et al (2017) Attenuation of insulin action by an allosteric insulin receptor antibody in healthy volunteers. J Clin Endocrinol Metab 102:3021–3028CrossRefPubMed

12.

Bedinger DH, Kieffer DA, Goldfine ID, Roell MK, Adams SH (2015) Acute treatment with XMetA activates hepatic insulin receptors and lowers blood glucose in normal mice. J Cell Biochem 116:2009–2019CrossRef

13.

Bhaskar V, Lau A, Goldfine ID et al (2013) XMetA, an allosteric monoclonal antibody to the insulin receptor, improves glycaemic control in mice with diet-induced obesity. Diabetes Obes Metab 15:272–275CrossRefPubMed

14.

Bhaskar V, Goldfine ID, Bedinger DH et al (2012) A fully human, allosteric monoclonal antibody that activates the insulin receptor and improves glycemic control. Diabetes 61:1263–1271CrossRefPubMedPubMedCentral

15.

Bezwada P, Zhao J, Der K et al (2016) A novel allosteric insulin receptor-activating antibody reduces hyperglycaemia without hypoglycaemia in diabetic cynomolgus monkeys. J Pharmacol Exp Ther 356:466–473CrossRefPubMed

16.

Soos MA, O’Brien RM, Brindle NP et al (1989) Monoclonal antibodies to the insulin receptor mimic metabolic effects of insulin but do not stimulate receptor autophosphorylation in transfected NIH 3T3 fibroblasts. Proc Natl Acad Sci U S A 86:5217–5221CrossRefPubMedPubMedCentral

17.

Soos MA, Siddle K, Baron MD et al (1986) Monoclonal antibodies reacting with multiple epitopes on the human insulin receptor. Biochem J 235:199–208CrossRefPubMedPubMedCentral

18.

Siddle K, Soos MA, O’Brien RM, Ganderton RH, Taylor R (1987) Monoclonal antibodies as probes of the structure and function of insulin receptors. Biochem Soc Trans 15:47–51CrossRefPubMed

19.

O’Brien RM, Soos MA, Siddle K (1987) Monoclonal antibodies to the insulin receptor stimulate the intrinsic tyrosine kinase activity by cross-linking receptor molecules. EMBO J 6:4003–4010PubMedCrossRef

20.

Taylor R, Soos MA, Wells A, Argyraki M, Siddle K (1987) Insulin-like and insulin-inhibitory effects of monoclonal antibodies for different epitopes on the human insulin receptor. Biochem J 242:123–129CrossRefPubMedPubMedCentral

21.

Shin KJ, Wall EA, Zavzavadjian JR et al (2006) A single lentiviral vector platform for microRNA-based conditional RNA interference and coordinated transgene expression. Proc Natl Acad Sci U S A 103:13759–13764CrossRefPubMedPubMedCentral

22.

Miki H, Yamauchi T, Suzuki RYO et al (2001) Essential role of insulin receptor substrate 1 (IRS-1) and IRS-2 in adipocyte differentiation. Mol Cell Biol 21:2521–2532CrossRefPubMedPubMedCentral

23.

Yamamoto N, Ueda M, Sato T et al (2011) Measurement of glucose uptake in cultured cells. Curr Protoc Pharmacol 1–22

24.

Krook A, Moller DE, Dib K, O’Rahilly S (1996) Two naturally occurring mutant insulin receptors phosphorylate insulin receptor substrate-1 (IRS-1) but fail to mediate the biological effects of insulin. Evidence that IRS-1 phosphorylation is not sufficient for normal insulin action. J Biol Chem 271:7134–7140CrossRefPubMed

25.

Krook A, Whitehead JP, Dobson SP et al (1997) Two naturally occurring insulin receptor tyrosine kinase domain mutants provide evidence that phosphoinositide 3-kinase activation alone is not sufficient for the mediation of insulin’s metabolic and mitogenic effects. J Biol Chem 272:30208–30214CrossRefPubMed

26.

Rouard M, Bass J, Grigorescu F et al (1999) Congenital insulin resistance associated with a conformational alteration in a conserved β-sheet in the insulin receptor L1 domain. J Biol Chem 274:18487–18491CrossRefPubMed

27.

Croll TI, Smith BJ, Margetts MB et al. (2016) Higher-resolution structure of the human insulin receptor ectodomain: multi-modal inclusion of the insert domain. Struct Des 1–8

28.

Menting JG, Whittaker J, Margetts MB et al (2013) How insulin engages its primary binding site on the insulin receptor. Nature 493:241–245CrossRefPubMedPubMedCentral

29.

Belfiore A, Frasca F, Pandini G, Sciacca L, Vigneri R (2009) Insulin receptor isoforms and insulin receptor/insulin-like growth factor receptor hybrids in physiology and disease. Endocr Rev 30:586–623CrossRefPubMed

30.

Rouard M, Macari F, Bouix O et al (1997) Identification of two novel insulin receptor mutations, Asp59Gly and Leu62Pro, in type A syndrome of extreme insulin resistance. Biochem Biophys Res Commun 234:764–768CrossRefPubMed

31.

Siddle K (2012) Molecular basis of signalling specificity of insulin and IGF receptors: neglected corners and recent advances. Front Endocrinol 3:1–24CrossRef

32.

Sciacca L, Cassarino MF, Genua M, Pandini G (2010) Insulin analogues differently activate insulin receptor isoforms and post-receptor signalling. Diabetologia 53:1743–1753CrossRefPubMed

33.

Hamer I, Foti M, Emkey R et al (2002) An arginine to cysteine 252 mutation in insulin receptors from a patient with severe insulin resistance inhibits receptor internalisation but preserves signalling events. Diabetologia 45:657–667CrossRefPubMed

34.

Zick Y, Rees-Jones RW, Taylor SI, Gorden P, Roth J (1984) The role of antireceptor antibodies in stimulating phosphorylation of the insulin receptor. J Biol Chem 259:4396–4400PubMed

35.

Sung CK, Maddux BA, Hawley DM, Goldfine ID (1989) Monoclonal antibodies mimic insulin activation of ribosomal protein S6 kinase without activation of insulin receptor tyrosine kinase. Studies in cells transfected with normal and mutant human insulin receptors. J Biol Chem 264:18951–18959PubMed

36.

Forsayeth JR, Caro JF, Sinha MK, Maddux BA, Goldfine ID (1987) Monoclonal antibodies to the human insulin receptor that activate glucose transport but not insulin receptor kinase activity. Proc Natl Acad Sci U S A 84:3448–3451CrossRefPubMedPubMedCentral

37.

Brindle NP, Tavare JM, Dickens M, Whittaker J, Siddle K (1990) Anti-insulin receptor monoclonal antibody-stimulated tyrosine phosphorylation in cells transfected with human insulin receptor cDNA. Biochem J 268:615–620CrossRefPubMedPubMedCentral

38.

Steele-Perkins G, Roth R (1990) Insulin-mimetic anti-insulin receptor monoclonal antibodies stimulate receptor kinase activity in intact cells. J Biol Chem 265:9458–9463PubMed

39.

Jensen M, Hansen B, De Meyts P, Schäffer L, Ursø B (2007) Activation of the insulin receptor by insulin and a synthetic peptide leads to divergent metabolic and mitogenic signalling and responses. J Biol Chem 282:35179–35186CrossRefPubMed

40.

Bedinger DH, Goldfine ID, Corbin JA, Roell MK, Adams SH (2015) Differential pathway coupling of the activated insulin receptor drives signalling selectivity by XMetA, an allosteric partial agonist antibody. J Pharmacol Exp Ther 353:35–43CrossRefPubMed

41.

Rakatzi I, Ramrath S, Ledwig D et al (2003) A novel insulin analogue with unique properties: LysB3, Glu B29 insulin induces prominent activation of insulin receptor substrate 2, but marginal phosphorylation of insulin receptor substrate 1. Diabetes 52:2227–2238CrossRefPubMed

42.

Ceresa BP, Kao AW, Santeler SR, Pessin JE (1998) Inhibition of clathrin-mediated endocytosis selectively attenuates specific insulin receptor signal transduction pathways. Mol Cell Biol 18:3862–3870CrossRefPubMedPubMedCentral

Title: Evaluation of anti-insulin receptor antibodies as potential novel therapies for human insulin receptoropathy using cell culture models
Authors: Gemma V. Brierley
Kenneth Siddle
Robert K. Semple
Publication date: 01-07-2018
Publisher: Springer Berlin Heidelberg
Published in: Diabetologia / Issue 7/2018
Print ISSN: 0012-186X
Electronic ISSN: 1432-0428
DOI: https://doi.org/10.1007/s00125-018-4606-2

ACC 2024 Congress Coverage

Heart Failure Video

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Evaluation of anti-insulin receptor antibodies as potential novel therapies for human insulin receptoropathy using cell culture models

Abstract

Aims/hypothesis

Methods

Results

Conclusions/interpretation

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Aims/hypothesis

Methods

Results

Conclusions/interpretation

Please log in to get access to this content

Other articles of this Issue 7/2018

Global burden of hypoglycaemia-related mortality in 109 countries, from 2000 to 2014: an analysis of death certificates

Up front

A proinflammatory CD4+ T cell phenotype in gestational diabetes mellitus

Circulating amino acids and the risk of macrovascular, microvascular and mortality outcomes in individuals with type 2 diabetes: results from the ADVANCE trial

Correction to: Clinically meaningful and lasting HbA1c improvement rarely occurs after 5 years of type 1 diabetes: an argument for early, targeted and aggressive intervention following diagnosis

A heterogeneous response of liver and skeletal muscle fat to the combination of a Paleolithic diet and exercise in obese individuals with type 2 diabetes: a randomised controlled trial

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page