Download - Insulin resistance 2013

Transcript
Page 1: Insulin resistance 2013

Sample & Assay Technologies

Insulin Resistance

Jesse Liang, Ph.D.

Page 2: Insulin resistance 2013

Sample & Assay Technologies Insulin

Insulin is produced by β cells of pancreas. Insulin causes cells to absorb glucose from the blood.

Two types of tissues are most strongly influenced by insulin: muscle cells (myocytes) and fat cells (adipocytes).

Page 3: Insulin resistance 2013

Sample & Assay Technologies Insulin signaling

When insulin knocks, GLUT4 opens the door. Measures of insulin resistance were lower in the patients with a PTEN mutation than in controls.The patients' insulin sensitivity could be explained by the presence of enhanced insulin signalingthrough the PI3K-AKT pathway, as evidenced by increased AKT phosphorylation.(New Engl J Med, Sep. 2012)

Page 4: Insulin resistance 2013

Sample & Assay Technologies Insulin resistance

Insulin resistance (insulin insensitivity, glucose intolerance) means that cells are less responsive to insulin.

Insulin stimulates glucose uptake in adipose tissue through the GLUT4 glucose transporter. GLUT4 expression or function regulates insulin sensitivity. Carbohydrate-responsive element-binding protein (ChREBP), a downstream target of GLUT4, is a major determinant of adipose tissue fatty acid synthesis and systemic insulin sensitivity (Nature, April 2012).

Page 5: Insulin resistance 2013

Sample & Assay TechnologiesCauses of insulin resistance

There are genetic insulin resistance and diet-induced insulin resistance.

Obesity is the most common cause of insulin resistance.

Overnutrition

inflammation oxidative stress lipid metabolism gastrointestinal microbiota(gut microbial influence)

Insulin resistance

Page 6: Insulin resistance 2013

Sample & Assay Technologies Inflammation

Adipose tissue inflammation

• Invasion of visceral adipose tissue by pro-inflammatory macrophages is considered to be a key event driving adipose-tissue inflammation.

• Visceral adipose tissue has a much greater negative metabolic effect than subcutaneous adipose tissue.

Liver inflammation

Hypothalamic inflammation

Central nervous system inflammation

Mechanisms:TNFα activates JNK and IκB Kinase (IKK), which pho IRS-1, hence inhibits insulin receptor signaling.

Page 7: Insulin resistance 2013

Sample & Assay Technologies Oxidative stress

Evidences:

Several cell culture models have shown that treatment of insulin-responsive cell lines with H2O2 causes a significant decrease in insulin sensitivity.

Induction of insulin resistance in cell culture with TNFα or glucocortocoidtreatment causes a significant induction of ROS. Insulin resistance in these models can be prevented by treatment with different antioxidant compounds.

Mechanisms:

1. Oxidative stress stimulates stress signaling such as JNK, that in turn cause inhibition of insulin signaling (mice without JNK1-signaling do not develop insulin resistance under dietary conditions that normally produce it).

2. ROS trigger the expression of MCP-1, activate NFκB, increase TNFα and IL-6 production, and promote macrophage infiltration.

Page 8: Insulin resistance 2013

Sample & Assay Technologies LipotoxicityLipotoxicity:Increased levels of circulating fatty acids and/or lipid accumulation inside thecells can lead to insulin resistance.

The origins and drivers of insulin resistance (Cell, Feb 2013)

Page 9: Insulin resistance 2013

Sample & Assay Technologies Microbial contribution to insulin resistance

Gnotobiotic (germ-free) mice are resistant to obesity induced by high fat diet.

Dysbiosis:In mouse models of obesity, the composition of the microbiota is altered.

A dysbiotic microbiota contributes to insulin resistance by increased energy harvest and the direct effect of altered bacterial metabolite production (e.g.,short-chain fatty acids and bile acid derivatives) on the liver and adipose tissue. In addition, gastrointestinal permeability caused by either intestinal inflammation or intestinal immune dysfunction results in penetration of bacteria or bacterial products (e.g., LPS, DNA), which can drive systemic inflammation.

Page 10: Insulin resistance 2013

Sample & Assay Technologies The major contributing or affected sites

Insulin resistance

Page 11: Insulin resistance 2013

Sample & Assay Technologies PPAR-γ is a major regulator of insulin sensitivity

Peroxisome proliferator-activated receptor (PPARγ) is the master regulator of adipogenesis, adipocyte differentiation and fat cell gene expression.

PPARγ is the functioning receptor for the thiazolidinedione (TZD) class of anti-diabetes drugs. These drugs are full classical agonists for this nuclear receptor.

PPARγ–FGF1 axis is critical for maintaining metabolic homeostasis and insulin sensitization (Nature, May 2012).

PPARγ is a crucial molecular orchestrator of visceral adipose tissue Treg cell accumulation, phenotype and function (Nature, June 2012).

Page 12: Insulin resistance 2013

Sample & Assay Technologies PPAR-γ is a major regulator of insulin sensitivity

PPAR-γ co-activator-1 α (PGC1-α) expression in muscle stimulates an increase in expression of FNDC5, a membrane protein that is cleaved and secreted as a newly identified hormone, irisin. Irisin acts on white adipose cells in culture and in vivo to stimulate UCP1 expression and a broad program of brown-fat-like development (Nature, Jan 2012).

Obesity induced in mice by high-fat feeding activates Cdk5 in adipose tissues. This results in phosphorylation of PPAR-γ at serine 273. This modification of PPAR-γ does not alter its adipogenic capacity, but leads to dysregulation of a large number of genes whose expression is altered in obesity, including a reduction in the expression of the insulin-sensitizing adipokine, adiponectin (Nature, July 2010).

Decreased levels of adiponectin and AdipoR1 in obesity may havecausal roles in mitochondrial dysfunction and insulin resistance. PGC1-α is a key regulator of mitochondrial content and function. Activities of PGC1-α can be modulated by AMPK and SIRT1 (Nature, April 2010).

Rb modulates the activity and the expression of Runx2 and PPAR-γ (Nature, August 2010).

Page 13: Insulin resistance 2013

Sample & Assay Technologies mTOR is another regulator of insulin resistance

mTOR stands for “the mechanistic target of rapamycin”

PPAR-γ and mTOR are 2 major regulators of insulin sensitivity.

mTORC1 and mTORC2 play distinct roles:mTORC1 inhibits insulin signaling through its substrate S6K1;mTORC2 increases insulin signaling through phosphorylation of Akt.

Page 14: Insulin resistance 2013

Sample & Assay Technologies A new energy-based concept

A new energy-based concept of insulin resistance, in which insulin resistance is a result of energy surplus in cells, starts to reinterpret literature for a unifying mechanism of insulin resistance. The energy surplus signal is mediated by ATP and sensed by AMPK signaling pathway. Decreasing ATP level by suppression of production or stimulation of utilization is a promising approach in the treatment of insulin resistance.

Page 15: Insulin resistance 2013

Sample & Assay Technologies AMPK is a crucial cellular energy sensor

AMPK stands for “AMP-activated protein kinase”.

AMPK monitors cellular energy status by sensing increases in the ratios of AMP/ATP and ADP/ATP.

AMPK is activated by low energy status (increased AMP/ADP: ATP) such as during exercise, and regulates metabolic process and energy homeostasis byswitching off ATP consuming pathways (fatty acid and cholesterol synthesis) and switching on ATP generating processes (glucose uptake and fatty acid oxidation).

Activation of AMPK promotes glucose uptake, fatty acid oxidation, mitochondrial biogenesis, and insulin sensitivity.

Page 16: Insulin resistance 2013

Sample & Assay Technologies Mitochondrial energy metabolism

http://www.sabiosciences.com/pathwaymagazine/minireview/mitchondrial_energy_metabolism.php

Page 17: Insulin resistance 2013

Sample & Assay Technologies Sirtuins controls acetylation state of histones and transcription factors

Originally, sirtuins were described as NAD-dependent type III HDACs.

The targets of SIRT1 are PGC1α, FOXO1, FOXO3, p53, Notch, NF-κB, HIF1α, LXR, FXR, SREBP1c and more.

Sirtuin activation prevents diet-induced obesity.

SIRT1 activation protects from diet-induced and genetic insulin resistance in mice. Mice lacking SIRT1 specifically in the liver have been shown to develop insulin resistance.

SIRT3 also has a role in insulin sensitization, as its absence may contribute to the development of insulin resistance in the muscle by increasing ROS production and impairing mitochondrial oxidation.

Page 18: Insulin resistance 2013

Sample & Assay Technologies Regulation of FOXO by class IIa HDACs

http://www.sabiosciences.com/pathwaymagazine/minireview/metabolichomeostasis.php

Page 19: Insulin resistance 2013

Sample & Assay Technologies miRNAs in insulin resistance

miR-103 and miR-107,

miR-143,

miR-223,

miR-33a and miR-33b,

miR-34a,

miR-378,

let-7

Page 20: Insulin resistance 2013

Sample & Assay Technologies Gene expression at mRNA level – PCR Arrays

• Insulin resistance• Insulin signaling• Glucose metabolism• Fatty acid metabolism• Mitochondrial energy metabolism• Fatty liver• Lipoprotein signaling and cholesterol metabolism• Oxidative stress• Glucocorticoid signaling • Autophagy• PPAR targets• mTOR signaling• Adipogenesis• Diabetes• Obesity• Hypertension• Atherosclerosis

http://www.sabiosciences.com/Metabolic_Diseases.php

Page 21: Insulin resistance 2013

Sample & Assay Technologies Gene expression at mRNA level – PCR Arrays

• Inflammatory cytokines and receptors• Chemokines and receptors• Common cytokines• Cytokines and chemokines• Antiviral response• Inflammatory response and autoimmunity• Toll-like receptors (TLRs)• Innate and adaptive immune response• Inflammasomes• IL-6/STAT3 signaling• T helper cell differentiation• Th1 and Th2 responses• Th17 response• Interferons and receptors• MAPK signaling• NFκB signaling• TGFβ / BMP signaling

http://www.sabiosciences.com/Cytokines_Inflammation.php

Page 22: Insulin resistance 2013

Sample & Assay Technologies PCR Array introduction

84 pathway-specific genes of interest

5 housekeeping genes

Genomic DNA contamination control

Reverse transcription controls (RTC) n=3

Positive PCR controls (PPC) n=3

Page 23: Insulin resistance 2013

Sample & Assay Technologies RT² Predictor PCR Arrays

Human Oxidative Stress Toxicity

Human Mitochondrial Energy Metabolism Toxicity

Human Hypoxia Signaling Pathway Activity

Human Unfolded Protein Response Toxicity

http://www.sabiosciences.com/pathwayactivity.php

Page 24: Insulin resistance 2013

Sample & Assay Technologies microRNA expression – microRNA PCR Arrays

• Liver miFinder

• Diabetes

• Cardiovascular diseases

• Immunopathology

• Inflammatory response and autoimmunity

• Serum and plasma miRNAs

http://www.sabiosciences.com/mirna_pcr_array.php

Page 25: Insulin resistance 2013

Sample & Assay Technologies We also provide services – simply send your samples

• RNA extraction

• DNA extraction

• Illumina chips

• All sorts of PCR Arrays

http://www.sabiosciences.com/servicecore.php

Page 26: Insulin resistance 2013

Sample & Assay Technologies Contact information

Jesse LiangEmail: [email protected]

Technical Support: 1-888-503-3187Email: [email protected]

Check Webinar Calendar:http://www.sabiosciences.com/seminarlist.php