Generation, validation and humanisation of a novel insulin resistant cell model

Lisa Logie, Antonio J. Ruiz-Alcaraz, Christopher J. Schofield, Hari S. Hundal, Giora Z. Feuerstein, Jeffrey D. Brady, Daniel Crowther, Anna M. Tommasi, Christal E. Grierson, Bridget Shepherd, Andrew D. Morris, Michael K. Hansen, Ewan Pearson, Calum Sutherland

    Research output: Contribution to journalArticle

    3 Citations (Scopus)

    Abstract

    Insulin resistance is a characteristic of type 2 diabetes and is a major independent risk factor for progression to the disease. In particular, insulin resistance associates with increased body fat and almost certainly contributes to the dramatic increase in risk of type 2 diabetes associated with obesity. Therefore, in order to design truly effective insulin sensitising agents, targeted at the mechanism of disease development, we aimed to generate an obesity-related insulin resistant cell model. Rat hepatoma cells were grown in the presence of serum isolated from obese rodents or obese human volunteers, and the insulin sensitivity of the cells monitored over time by measuring a well-characterised insulin regulated gene promoter. Higher insulin concentrations were required to fully repress the gene in the cells grown in obese rodent serum compared with those grown in serum from lean rodents (almost a 10-fold shift in insulin sensitivity). This was reversed by restoration of normal growth medium, while the insulin resistance was prevented by pioglitazone or metformin. Meanwhile, growth of cells in serum collected from obese human volunteers with diabetes also reduced the insulin sensitivity of the rat cells. No clinical marker predicted the degree of insulin resistance that was generated by the human serum. We have developed a novel insulin resistant cell model for the study of the molecular development of obesity-linked insulin resistance, screen for compounds to overcome obesity-related insulin resistance and potentially search for novel serum biomarkers of insulin resistance. (C) 2010 Elsevier Inc. All rights reserved.

    Original languageEnglish
    Pages (from-to)1042-1049
    Number of pages8
    JournalBiochemical Pharmacology
    Volume80
    Issue number7
    DOIs
    Publication statusPublished - 1 Oct 2010

    Keywords

    • Insulin sensitivity
    • Liver
    • Pepck
    • Diabetes
    • Obesity
    • GLYCOGEN-SYNTHASE KINASE-3
    • PHOSPHATIDYLINOSITOL 3-KINASE ACTIVITY
    • RECEPTOR SUBSTRATE-1 PHOSPHORYLATION
    • CARBOXYKINASE GENE-EXPRESSION
    • PROTEIN-KINASE
    • SKELETAL-MUSCLE
    • GLUCOSE-METABOLISM
    • RESPONSE ELEMENT
    • IN-VITRO
    • OBESITY

    Cite this

    Logie, Lisa ; Ruiz-Alcaraz, Antonio J. ; Schofield, Christopher J. ; Hundal, Hari S. ; Feuerstein, Giora Z. ; Brady, Jeffrey D. ; Crowther, Daniel ; Tommasi, Anna M. ; Grierson, Christal E. ; Shepherd, Bridget ; Morris, Andrew D. ; Hansen, Michael K. ; Pearson, Ewan ; Sutherland, Calum. / Generation, validation and humanisation of a novel insulin resistant cell model. In: Biochemical Pharmacology. 2010 ; Vol. 80, No. 7. pp. 1042-1049.
    @article{6430a21c9b944f72acea19e6449aa989,
    title = "Generation, validation and humanisation of a novel insulin resistant cell model",
    abstract = "Insulin resistance is a characteristic of type 2 diabetes and is a major independent risk factor for progression to the disease. In particular, insulin resistance associates with increased body fat and almost certainly contributes to the dramatic increase in risk of type 2 diabetes associated with obesity. Therefore, in order to design truly effective insulin sensitising agents, targeted at the mechanism of disease development, we aimed to generate an obesity-related insulin resistant cell model. Rat hepatoma cells were grown in the presence of serum isolated from obese rodents or obese human volunteers, and the insulin sensitivity of the cells monitored over time by measuring a well-characterised insulin regulated gene promoter. Higher insulin concentrations were required to fully repress the gene in the cells grown in obese rodent serum compared with those grown in serum from lean rodents (almost a 10-fold shift in insulin sensitivity). This was reversed by restoration of normal growth medium, while the insulin resistance was prevented by pioglitazone or metformin. Meanwhile, growth of cells in serum collected from obese human volunteers with diabetes also reduced the insulin sensitivity of the rat cells. No clinical marker predicted the degree of insulin resistance that was generated by the human serum. We have developed a novel insulin resistant cell model for the study of the molecular development of obesity-linked insulin resistance, screen for compounds to overcome obesity-related insulin resistance and potentially search for novel serum biomarkers of insulin resistance. (C) 2010 Elsevier Inc. All rights reserved.",
    keywords = "Insulin sensitivity, Liver, Pepck, Diabetes, Obesity, GLYCOGEN-SYNTHASE KINASE-3, PHOSPHATIDYLINOSITOL 3-KINASE ACTIVITY, RECEPTOR SUBSTRATE-1 PHOSPHORYLATION, CARBOXYKINASE GENE-EXPRESSION, PROTEIN-KINASE, SKELETAL-MUSCLE, GLUCOSE-METABOLISM, RESPONSE ELEMENT, IN-VITRO, OBESITY",
    author = "Lisa Logie and Ruiz-Alcaraz, {Antonio J.} and Schofield, {Christopher J.} and Hundal, {Hari S.} and Feuerstein, {Giora Z.} and Brady, {Jeffrey D.} and Daniel Crowther and Tommasi, {Anna M.} and Grierson, {Christal E.} and Bridget Shepherd and Morris, {Andrew D.} and Hansen, {Michael K.} and Ewan Pearson and Calum Sutherland",
    year = "2010",
    month = "10",
    day = "1",
    doi = "10.1016/j.bcp.2010.06.011",
    language = "English",
    volume = "80",
    pages = "1042--1049",
    journal = "Biochemical Pharmacology",
    issn = "0006-2952",
    publisher = "Elsevier",
    number = "7",

    }

    Logie, L, Ruiz-Alcaraz, AJ, Schofield, CJ, Hundal, HS, Feuerstein, GZ, Brady, JD, Crowther, D, Tommasi, AM, Grierson, CE, Shepherd, B, Morris, AD, Hansen, MK, Pearson, E & Sutherland, C 2010, 'Generation, validation and humanisation of a novel insulin resistant cell model', Biochemical Pharmacology, vol. 80, no. 7, pp. 1042-1049. https://doi.org/10.1016/j.bcp.2010.06.011

    Generation, validation and humanisation of a novel insulin resistant cell model. / Logie, Lisa; Ruiz-Alcaraz, Antonio J.; Schofield, Christopher J.; Hundal, Hari S.; Feuerstein, Giora Z.; Brady, Jeffrey D.; Crowther, Daniel; Tommasi, Anna M.; Grierson, Christal E.; Shepherd, Bridget; Morris, Andrew D.; Hansen, Michael K.; Pearson, Ewan; Sutherland, Calum.

    In: Biochemical Pharmacology, Vol. 80, No. 7, 01.10.2010, p. 1042-1049.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Generation, validation and humanisation of a novel insulin resistant cell model

    AU - Logie, Lisa

    AU - Ruiz-Alcaraz, Antonio J.

    AU - Schofield, Christopher J.

    AU - Hundal, Hari S.

    AU - Feuerstein, Giora Z.

    AU - Brady, Jeffrey D.

    AU - Crowther, Daniel

    AU - Tommasi, Anna M.

    AU - Grierson, Christal E.

    AU - Shepherd, Bridget

    AU - Morris, Andrew D.

    AU - Hansen, Michael K.

    AU - Pearson, Ewan

    AU - Sutherland, Calum

    PY - 2010/10/1

    Y1 - 2010/10/1

    N2 - Insulin resistance is a characteristic of type 2 diabetes and is a major independent risk factor for progression to the disease. In particular, insulin resistance associates with increased body fat and almost certainly contributes to the dramatic increase in risk of type 2 diabetes associated with obesity. Therefore, in order to design truly effective insulin sensitising agents, targeted at the mechanism of disease development, we aimed to generate an obesity-related insulin resistant cell model. Rat hepatoma cells were grown in the presence of serum isolated from obese rodents or obese human volunteers, and the insulin sensitivity of the cells monitored over time by measuring a well-characterised insulin regulated gene promoter. Higher insulin concentrations were required to fully repress the gene in the cells grown in obese rodent serum compared with those grown in serum from lean rodents (almost a 10-fold shift in insulin sensitivity). This was reversed by restoration of normal growth medium, while the insulin resistance was prevented by pioglitazone or metformin. Meanwhile, growth of cells in serum collected from obese human volunteers with diabetes also reduced the insulin sensitivity of the rat cells. No clinical marker predicted the degree of insulin resistance that was generated by the human serum. We have developed a novel insulin resistant cell model for the study of the molecular development of obesity-linked insulin resistance, screen for compounds to overcome obesity-related insulin resistance and potentially search for novel serum biomarkers of insulin resistance. (C) 2010 Elsevier Inc. All rights reserved.

    AB - Insulin resistance is a characteristic of type 2 diabetes and is a major independent risk factor for progression to the disease. In particular, insulin resistance associates with increased body fat and almost certainly contributes to the dramatic increase in risk of type 2 diabetes associated with obesity. Therefore, in order to design truly effective insulin sensitising agents, targeted at the mechanism of disease development, we aimed to generate an obesity-related insulin resistant cell model. Rat hepatoma cells were grown in the presence of serum isolated from obese rodents or obese human volunteers, and the insulin sensitivity of the cells monitored over time by measuring a well-characterised insulin regulated gene promoter. Higher insulin concentrations were required to fully repress the gene in the cells grown in obese rodent serum compared with those grown in serum from lean rodents (almost a 10-fold shift in insulin sensitivity). This was reversed by restoration of normal growth medium, while the insulin resistance was prevented by pioglitazone or metformin. Meanwhile, growth of cells in serum collected from obese human volunteers with diabetes also reduced the insulin sensitivity of the rat cells. No clinical marker predicted the degree of insulin resistance that was generated by the human serum. We have developed a novel insulin resistant cell model for the study of the molecular development of obesity-linked insulin resistance, screen for compounds to overcome obesity-related insulin resistance and potentially search for novel serum biomarkers of insulin resistance. (C) 2010 Elsevier Inc. All rights reserved.

    KW - Insulin sensitivity

    KW - Liver

    KW - Pepck

    KW - Diabetes

    KW - Obesity

    KW - GLYCOGEN-SYNTHASE KINASE-3

    KW - PHOSPHATIDYLINOSITOL 3-KINASE ACTIVITY

    KW - RECEPTOR SUBSTRATE-1 PHOSPHORYLATION

    KW - CARBOXYKINASE GENE-EXPRESSION

    KW - PROTEIN-KINASE

    KW - SKELETAL-MUSCLE

    KW - GLUCOSE-METABOLISM

    KW - RESPONSE ELEMENT

    KW - IN-VITRO

    KW - OBESITY

    U2 - 10.1016/j.bcp.2010.06.011

    DO - 10.1016/j.bcp.2010.06.011

    M3 - Article

    C2 - 20599791

    VL - 80

    SP - 1042

    EP - 1049

    JO - Biochemical Pharmacology

    JF - Biochemical Pharmacology

    SN - 0006-2952

    IS - 7

    ER -