PEGylation of lysine residues reduces the pro-migratory activity of IGF-I.

BACKGROUND The insulin-like growth factor (IGF) system is composed of ligands and receptors which regulate cell proliferation, survival, differentiation and migration. Some of these functions involve regulation by the extracellular milieu, including binding proteins and other extracellular matrix proteins. However, the functions and exact nature of these interactions remain incomplete. METHODS IGF-I variants PEGylated at lysines K27, K65 and K68, were assessed for binding to IGFBPs using BIAcore, and for phosphorylation of the IGF-IR. Furthermore, functional consequences of PEGylation were investigated using cell viability and migration assays. In addition, downstream signaling pathways were analyzed using phospho-AKT and phospho-ERK1/2 assays. RESULTS IGF-I PEGylated at lysines 27 (PEG-K27), 65 (PEG-K65) or 68 (PEG-K68) was employed. Receptor phosphorylation was similarly reduced 2-fold with PEG-K65 and PEG-K68 in 3T3 fibroblasts and MCF-7 breast cancer cells, whereas PEG-K27 showed a more than 10- and 3-fold lower activation for 3T3 and MCF-7 cells, respectively. In addition, all PEG-IGF-I variants had a 10-fold reduced association rate to IGF binding proteins (IGFBPs). Functionally, all PEG variants lost their ability to induce cell migration in the presence of IGFBP-3/vitronectin (VN) complexes, whereas cell viability was fully preserved. Analysis of downstream signaling revealed that AKT was preferentially affected upon treatment with PEG-IGF-I variants whereas MAPK signaling was unaffected by PEGylation. CONCLUSION PEGylation of IGF-I has an impact on cell migration but not on cell viability. GENERAL SIGNIFICANCE PEG-IGF-I may differentially modulate IGF-I mediated functions that are dependent on receptor interaction as well as key extracellular proteins such as VN and IGFBPs.

• Publication year

  2013

• Venue

  Biochimica et Biophysica Acta

• Publication date

  2013-06-01

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1016/j.bbagen.2013.06.012PMID 23800578
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Functional improvement in mouse models of familial amyotrophic lateral sclerosis by PEGylated insulin-like growth factor I treatment depends on disease severity

  2012cited by this paper
• Metabolic actions of insulin-like growth factor-I in normal physiology and diabetes.

  2012cited by this paper
• The insulin and insulin-like growth factor receptor family in neoplasia: an update

  2012cited by this paper
• Therapeutic potential of PEGylated insulin-like growth factor I for skeletal muscle disease evaluated in two murine models of muscular dystrophy.

  2012cited by this paper
• Clinical development of insulin-like growth factor receptor—1 (IGF-1R) inhibitors: At the crossroad?

  2012cited by this paper
• Targeting the insulin-like growth factor 1 receptor (IGF1R) signaling pathway for cancer therapy

  2012cited by this paper
• Signalling pathways of insulin-like growth factors (IGFs) and IGF binding protein-3

  2011cited by this paper
• Therapeutic effects of PEGylated insulin-like growth factor I in the pmn mouse model of motoneuron disease.

  2011cited by this paper
• Transglutaminases and receptor tyrosine kinases

  2011cited by this paper
• PEGylation enhances the therapeutic potential for insulin-like growth factor I in central nervous system disorders.

  2011cited by this paper
• Separation of Fast from Slow Anabolism by Site-specific PEGylation of Insulin-like Growth Factor I (IGF-I)*

  2011cited by this paper
• Insulin-like growth factor-I:vitronectin complex-induced changes in gene expression effect breast cell survival and migration.

  2011cited by this paper
• Mechanistic investigations into interactions between IGF-I and IGFBPs and their impact on facilitating cell migration on vitronectin

  2010cited by this paper
• Abstract S1-4: A Randomized, Double-Blind, Placebo-Controlled, Phase 2 Study of AMG 479 With Exemestane (E) or Fulvestrant (F) in Postmenopausal Women With Hormone-Receptor Positive (HR+) Metastatic (M) or Locally Advanced (LA) Breast Cancer (BC)

  2010cited by this paper
• Functional and phenotypic characterization of human keratinocytes expanded in microcarrier culture.

  2009cited by this paper
• The IGF-1/IGF-1R signaling axis in the skin: a new role for the dermis in aging-associated skin cancer

  2009cited by this paper
• How does suppression of IGF-1 signaling by DNA damage affect aging and longevity?

  2008cited by this paper
• Vitronectin: growth factor complexes hold potential as a wound therapy approach.

  2008cited by this paper
• Substrate-bound insulin-like growth factor (IGF)-I-IGF binding protein-vitronectin-stimulated breast cell migration is enhanced by coactivation of the phosphatidylinositide 3-Kinase/AKT pathway by alphav-integrins and the IGF-I receptor.

  2008cited by this paper
• Development of Defined Media for the Serum-Free Expansion of Primary Keratinocytes and Human Embryonic Stem Cells

  2008cited by this paper
• The emerging role of the insulin-like growth factor pathway as a therapeutic target in cancer.

  2008cited by this paper
• UVB-induced senescence in human keratinocytes requires a functional insulin-like growth factor-1 receptor and p53.

  2008cited by this paper
• Increased matrix metalloproteinase‐9 (MMP‐9) activity observed in chronic wound fluid is related to the clinical severity of the ulcer

  2008cited by this paper
• Constitutively Active Type I Insulin-Like Growth Factor Receptor Causes Transformation and Xenograft Growth of Immortalized Mammary Epithelial Cells and Is Accompanied by an Epithelial-to-Mesenchymal Transition Mediated by NF-κB and Snail

  2007cited by this paper
• Efficacy of the anti-insulin like growth factor I receptor (IGF-IR) antibody CP-751871 in combination with paclitaxel and carboplatin as first-line treatment for advanced non-small cell lung cancer (NSCLC)

  2007cited by this paper
• Divergent Roles for IRS-1 and IRS-2 in Breast Cancer Metastasis

  2007cited by this paper
• Complex networks orchestrate epithelial–mesenchymal transitions

  2006cited by this paper
• Molecular interactions of the IGF system.

  2005cited by this paper
• Recombinant Human Insulin-Like Growth Factor I (IGF-I): Risks and Benefits of Normalizing Blood IGF-I Concentrations

  2005cited by this paper
• Responses of keratinocytes to substrate-bound vitronectin: growth factor complexes.

  2005cited by this paper
• IGF-I and Longevity

  2004cited by this paper
• Focal adhesion regulation of cell behavior.

  2004cited by this paper
• Minireview: Integral membrane proteins that function coordinately with the insulin-like growth factor I receptor to regulate intracellular signaling.

  2003cited by this paper
• The muscular dystrophies.

  2002cited by this paper
• Insulin-like Growth Factor-1-mediated AKT Activation Postpones the Onset of Ultraviolet B-induced Apoptosis, Providing More Time for Cyclobutane Thymine Dimer Removal in Primary Human Keratinocytes*

  2002cited by this paper
• Lack of insulin‐like growth factor 1 (IGF1) in the basal keratinocyte layer of diabetic skin and diabetic foot ulcers

  2000cited by this paper
• Activation of the insulin‐like growth factor‐1 receptor promotes the survival of human keratinocytes following ultraviolet B irradiation

  1999cited by this paper
• Insulin-like Growth Factors as Regulators of Cell Motility Signaling Mechanisms.

  1997cited by this paper
• A branched monomethoxypoly(ethylene glycol) for protein modification.

  1995cited by this paper
• Modification of CD4 immunoadhesin with monomethoxypoly(ethylene glycol) aldehyde via reductive alkylation.

  1994cited by this paper
• Insulin-like growth factor I preserves host lean tissue mass in cancer cachexia.

  1992cited by this paper
• Chemical modification of recombinant human granulocyte colony-stimulating factor by polyethylene glycol increases its biological activity in vivo.

  1992cited by this paper
• Use of an aqueous soluble tetrazolium/formazan assay for cell growth assays in culture.

  1991cited by this paper
• A phase I study including pharmacokinetics of polyethylene glycol conjugated interleukin‐2

  1991cited by this paper
• Mutants of human insulin-like growth factor II: expression and characterization of analogs with a substitution of TYR27 and/or a deletion of residues 62-67.

  1991cited by this paper
• The roles of tyrosines 24, 31, and 60 in the high affinity binding of insulin-like growth factor-I to the type 1 insulin-like growth factor receptor.

  1990cited by this paper
• Immunogenicity of recombinant IL-2 modified by covalent attachment of polyethylene glycol.

  1990cited by this paper
• Mutants of human insulin-like growth factor I with reduced affinity for the type 1 insulin-like growth factor receptor.

  1988cited by this paper
• Treatment of adenosine deaminase deficiency with polyethylene glycol-modified adenosine deaminase.

  1987cited by this paper

• Improved Intranasal Retentivity and Transnasal Absorption Enhancement by PEGylated Poly-l-ornithine

  2018cites this paper
• Bioresponsive release of insulin‐like growth factor‐I from its PEGylated conjugate

  2018cites this paper
• Modification with polysialic acid–PEG copolymer as a new method for improving the therapeutic efficacy of proteins

  2016cites this paper
• Drug delivery of Insulin-like growth factor I.

  2015cites this paper
• Cell-based assays for IGF-I bioactivity measurement: overview, limitations and current trends

  2014cites this paper
• The Complexity of the IGF1 Gene Splicing, Posttranslational Modification and Bioactivity

  2014cites this paper