Team:BGU Israel/Project/Hormone Workshop

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        <p><b>The Problem: </b> Abnormal aggregation of fatty acid in non-adipose  tissues, particularly skeletal muscles and liver </p>
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        <p><b>The Goal: </b>Reduce insulin resistance and improve the metabolism  of metabolic syndrome patients</p>
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         <h3 style="border-bottom:dashed;border-color:#000000">Mechanism  <a href="#" onclick="goToByScroll('test'); return false;" class="right"></a></h3>
         <h3 style="border-bottom:dashed;border-color:#000000">Mechanism  <a href="#" onclick="goToByScroll('test'); return false;" class="right"></a></h3>
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The most active form of the Adiponectin is a high molecular weight complex. The synthesis of the protein subunit and an enzyme for disulfide bonds should improve the levels of the active form of the hormone.
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       <h3 style="border-bottom:dashed;border-color:#000000">Background </h3>
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         <p>Adiponectin is an adipocyte derived protein hormone showing insulin- sensitizing effect and important role in lipid and glucose metabolism regulation. Hypoadiponectemia (low levels of serum Adiponectin) is highly related with occurrence of abdominal obesity, insulin resistance, type 2 diabetes, and the metabolic syndrome. Adiponectin forms multimeric complexes that combine via its collagen domain to create three major oligomeric forms: a low molecular weight (LMW) trimer, a middle - molecular weight (MMW) hexamer, and high molecular weight (HMW) 12-18- mer Adiponectin. The form showing the most <div id="test"></div> relevant properties considering insulin sensitivity and protection from diabetes is the HMW form of the hormone (1)  (figure 1.).</p>
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         <p>Adiponectin is an adipocyte derived protein hormone showing insulin - sensitizing effect and has an important role in lipid and glucose metabolism regulation. Hypoadiponectemia (low levels of serum Adiponectin) is highly related with occurrence of abdominal obesity, insulin resistance, type 2 diabetes, and the metabolic syndrome. Adiponectin forms multimeric complexes that combine via its collagen domain to create three major oligomeric forms: a low molecular weight (LMW) trimer, a middle - molecular weight (MMW) hexamer, and high molecular weight (HMW) 12-18- mer Adiponectin. The form showing the most relevant properties considering insulin sensitivity and protection from diabetes is the HMW form of the hormone (Kadowaki et al., 2006).      
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<p align="center"><img src="https://static.igem.org/mediawiki/2014/4/41/BGU14_ANI_ADIPO.png" height="250"></p>
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  <img src="https://static.igem.org/mediawiki/2014/8/83/BGU14figure1-adiponectin.png" /> </p>
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  <img src="https://static.igem.org/mediawiki/2014/1/12/BGU14adipo_figure_1.png"  width="330"/> </p>
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  <p align="center" style="font-size:14px; font-weight:bold;">Figure 1. Adiponectin oligomeric forms  
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  <p align="center" style="font-size:14px; font-weight:bold;">Adiponectin oligomeric forms  
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       <h3 align="left" style="border-bottom:dashed;border-color:#000000">Mechanism </h3>
       <h3 align="left" style="border-bottom:dashed;border-color:#000000">Mechanism </h3>
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                 <p>When one is diagnosed with the metabolic syndrome's symptoms and shows low levels of serum Adiponectin, an efficient treatment could be a pill or a shot containing the active HMW form of Adiponectin hormone. However, there is no known efficient synthetic method to produce the desired multimer structure of the hormone. It has been shown that overexpression of the enzyme DsbA-L (disulfide-bond A oxidoreductase - like), promotes Adiponectin multimerization, while suppressing its expression by RNAi markedly and selectively reduced Adiponectin levels and secretion in adipocytes (2).<br>
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                 <p>When one is diagnosed with the metabolic syndrome's symptoms and shows low levels of serum Adiponectin, an efficient treatment could be a pill or a shot containing the active HMW form of Adiponectin hormone. However, there is no known efficient synthetic method to produce the desired multimer structure of the hormone. It has been shown that overexpression of the enzyme DsbA-L (disulfide-bond A oxidoreductase - like), promotes Adiponectin multimerization, while suppressing its expression by RNAi markedly and selectively reduced Adiponectin levels and secretion in adipocytes (Liu et al., 2008).<br>
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                In order to synthesize the HMW form of Adiponectin, we intend to introduce overexpression of both Adiponectin and DsbA-L in our model cell line HepG2 (human hepatocellular carcinoma). This is expected to produces high levels of HMW Adiponectin (figure  2.) while relying on the natural hormone secreting ability of hepatocytes and excretion tags on the hormone itself. </p>
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          In order to synthesize the HMW form of Adiponectin, we intend to introduce overexpression of both Adiponectin and DsbA-L in our model cell line HepG2 (human hepatocellular carcinoma). This is expected to produces high levels of HMW Adiponectin while relying on the natural hormone secreting ability of hepatocytes and excretion tags on the hormone itself.<br>
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<p>In the future, we intend to attach the hormone to an inducible system to allow for more control on the proteins synthesis. </p>
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                In the future, we intend to attach the hormone to an inducible system to allow for more control on the proteins synthesis. </p>
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           <p align="center" style="border: thin solid; border-radius: 8px; background: #5FA6B8; font-style: normal; font-weight: 700; font-size: 24px; font-family: Allan, 'Hoefler Text', 'Liberation Serif', Times, 'Times New Roman', serif;"> Click on the picture to check out the machanism <img src="https://static.igem.org/mediawiki/2014/f/f5/BGU14finger-point2.png" width="26px"/></p>
           <p align="center" style="border: thin solid; border-radius: 8px; background: #5FA6B8; font-style: normal; font-weight: 700; font-size: 24px; font-family: Allan, 'Hoefler Text', 'Liberation Serif', Times, 'Times New Roman', serif;"> Click on the picture to check out the machanism <img src="https://static.igem.org/mediawiki/2014/f/f5/BGU14finger-point2.png" width="26px"/></p>
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        <img src="https://static.igem.org/mediawiki/2014/7/75/BGU14adipo_figure_2_a.png" onclick="change_pic_hormone()" width="400" style="border: thin solid; cursor: pointer;" />
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          <p align="center" style="font-size:14px; font-weight:bold; line-height:normal">Adiponectin is overexpressed</p>
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        <img src="https://static.igem.org/mediawiki/2014/c/cb/BGU14adipo_figure_2_b.png" onclick="change_pic_hormone()" width="400" style="border: thin solid; cursor: pointer;" />
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        <p align="center" style="font-size:14px; font-weight:bold; line-height:normal">DsbA-L is overexpressed</p>
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        <img  src="https://static.igem.org/mediawiki/2014/1/1b/BGU14adipo_figure_2_c.png" onclick="change_pic_hormone()" width="400" style="border: thin solid; cursor: pointer;" />
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         <p align="center" style="font-size:14px; font-weight:bold; line-height:normal">Figure 2 –  Need to put content</p>
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         <p align="center" style="font-size:14px; font-weight:bold; line-height:normal">DsbA-L promotes Adiponectin multimerization leading to high levels of HMW Adiponectin</p>
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       <h3 style="border-bottom:dashed;border-color:#000000">References </h3>
       <h3 style="border-bottom:dashed;border-color:#000000">References </h3>
      
      
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       <p>Kadowaki, T. et al. 2006 Adiponectin and adiponectin receptors in insulin resistance, diabetes, and the metabolic syndrome. J Clin Invest 116:1784–1792 </p>
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       <p>Kadowaki, T., Yamauchi, T., Kubota, N., Hara, K., Ueki, K.,  &amp; Tobe, K. (2006). Review series Adiponectin and adiponectin receptors in insulin resistance , diabetes , and the metabolic syndrome. <em>J Clin Invest.</em>, <em>116</em>(7), 1784–1792. doi:10.1172/JCI29126.1784<br><br>
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      <p dir="LTR">Liu, M.&nbsp;<em>et al</em>.&nbsp;2008.&nbsp;A disulfide-bond A oxidoreductase-like protein (DsbA-L) regulates adiponectin multimerization.&nbsp;<em>Proc. Natl. Acad. Sci. USA</em>&nbsp;<strong>105</strong>:&nbsp;18302–18307.</p>
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        Liu,  M., Zhou, L., Xu, A., Lam, K. S. L., Wetzel, M. D., Xiang, R., Liu, F. (2008). A disulfide-bond A oxidoreductase-like protein (DsbA-L) regulates adiponectin multimerization. <em>Proceedings of the National Academy of Sciences of the  United States of America</em>, <em>105</em>(47), 18302–7.  doi:10.1073/pnas.0806341105</p>
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Latest revision as of 00:00, 18 October 2014

Backgroud

The Problem: Abnormal aggregation of fatty acid in non-adipose tissues, particularly skeletal muscles and liver

The Goal: Reduce insulin resistance and improve the metabolism of metabolic syndrome patients

Mechanism


The most active form of the Adiponectin is a high molecular weight complex. The synthesis of the protein subunit and an enzyme for disulfide bonds should improve the levels of the active form of the hormone.

Background

Adiponectin is an adipocyte derived protein hormone showing insulin - sensitizing effect and has an important role in lipid and glucose metabolism regulation. Hypoadiponectemia (low levels of serum Adiponectin) is highly related with occurrence of abdominal obesity, insulin resistance, type 2 diabetes, and the metabolic syndrome. Adiponectin forms multimeric complexes that combine via its collagen domain to create three major oligomeric forms: a low molecular weight (LMW) trimer, a middle - molecular weight (MMW) hexamer, and high molecular weight (HMW) 12-18- mer Adiponectin. The form showing the most relevant properties considering insulin sensitivity and protection from diabetes is the HMW form of the hormone (Kadowaki et al., 2006).

Adiponectin oligomeric forms

Mechanism

When one is diagnosed with the metabolic syndrome's symptoms and shows low levels of serum Adiponectin, an efficient treatment could be a pill or a shot containing the active HMW form of Adiponectin hormone. However, there is no known efficient synthetic method to produce the desired multimer structure of the hormone. It has been shown that overexpression of the enzyme DsbA-L (disulfide-bond A oxidoreductase - like), promotes Adiponectin multimerization, while suppressing its expression by RNAi markedly and selectively reduced Adiponectin levels and secretion in adipocytes (Liu et al., 2008).
In order to synthesize the HMW form of Adiponectin, we intend to introduce overexpression of both Adiponectin and DsbA-L in our model cell line HepG2 (human hepatocellular carcinoma). This is expected to produces high levels of HMW Adiponectin while relying on the natural hormone secreting ability of hepatocytes and excretion tags on the hormone itself.
In the future, we intend to attach the hormone to an inducible system to allow for more control on the proteins synthesis.

Click on the picture to check out the machanism

Adiponectin is overexpressed

References

Kadowaki, T., Yamauchi, T., Kubota, N., Hara, K., Ueki, K., & Tobe, K. (2006). Review series Adiponectin and adiponectin receptors in insulin resistance , diabetes , and the metabolic syndrome. J Clin Invest., 116(7), 1784–1792. doi:10.1172/JCI29126.1784

Liu, M., Zhou, L., Xu, A., Lam, K. S. L., Wetzel, M. D., Xiang, R., Liu, F. (2008). A disulfide-bond A oxidoreductase-like protein (DsbA-L) regulates adiponectin multimerization. Proceedings of the National Academy of Sciences of the United States of America, 105(47), 18302–7. doi:10.1073/pnas.0806341105