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Identification of a novel function of adipocyte plasma membrane-associated protein (APMAP) in gestational diabetes mellitus by proteomic analysis of omental adipose tissue Yuhang Ma, Jing Gao, Jiajing Yin, Liping Gu, Xing Liu, Su Chen, Qianfang Huang, Huifang Lu, Yuemin Yang, Hu Zhou, Yufan Wang, and Yongde Peng J. Proteome Res., Just Accepted Manuscript • DOI: 10.1021/acs.jproteome.5b01030 • Publication Date (Web): 15 Jan 2016 Downloaded from http://pubs.acs.org on January 24, 2016
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Journal of Proteome Research is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.
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Journal of Proteome Research
Identification
of
a
novel
function
of
adipocyte
plasma
membrane-associated protein (APMAP) in gestational diabetes mellitus by proteomic analysis of omental adipose tissue Yuhang Ma1, Jing Gao3, Jiajing Yin1, Liping Gu1, Xing Liu3, Su Chen1, Qianfang Huang1, Huifang Lu2, Yuemin Yang2, Hu Zhou3*, Yufan Wang1* and Yongde Peng1*
1
Department of Endocrinology and Metabolism, 2Department of Obstetrics and
Gynecology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China, 200080. 3 Department of Analytical Chemistry and CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China, 201203
* Correspondence: Hu Zhou; E-mail:
[email protected], Tel: +86-021-50806706, Department of Analytical Chemistry and CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road, Shanghai, China 201203
Yongde Peng, E-mail:
[email protected], Tel: +86-21-63240090-3732, Department of Endocrinology and Metabolism, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 100 Haining Road, Shanghai, China, 200080.
Yufan
Wang,
E-mail:
[email protected],
Tel:
+86-21-63240090-8589,
Department of Endocrinology and Metabolism, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 650 Xin Songjiang Road, Shanghai, China, 201620.
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Abstract Gestational diabetes mellitus (GDM) is considered as an early stage of type 2 diabetes mellitus. In this study, we compared demographic and clinical data between six GDM subjects and six normal glucose tolerance (NGT; healthy controls) subjects and found that homeostasis model of assessment for insulin resistence index (HOMA-IR) increased in GDM. Many previous studies demonstrated that omental adipose tissue dysfunction could induce insulin resistance. Thus, to investigate the cause of insulin resistance
in
GDM,
label-free
proteomics
was
used
to
identify
differentially expressed proteins in omental adipose tissues from GDM and NGT subjects. A total of 3528 proteins were identified, including 66 significantly changed proteins. Adipocyte plasma membrane-associated protein (APMAP, a.k.a. C20orf3), one of the differentially expressed proteins, was down regulated in GDM omental adipose tissues. Furthermore, mature 3T3-L1 adipocytes were used to simulate omental adipocytes. The inhibition of APMAP expression by RNAi impaired insulin signaling and activated NFκB signaling in these adipocytes. Our study revealed that the down-regulation of APMAP in omental adipose tissue may play an important role in insulin resistance in the pathophysiology of GDM.
Keywords: Gestational diabetes mellitus; Label-free proteomic analysis; Insulin resistance; Inflammation
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Journal of Proteome Research
Introduction Gestational diabetes mellitus (GDM) is a special type of diabetes; most patients return to normal blood glucose levels after delivery 1. More importantly, women with GDM have an increased risk of postpartum diabetes compared to women with normal glucose tolerance during pregnancy
2
. Recently, GDM has been considered as an
early stage of type 2 diabetes mellitus (T2DM) 3. It is generally recognized that the pathophysiological mechanism of GDM is insulin resistance, rather than β cell dysfunction
4, 5
. However, the mechanism of insulin resistance in GDM remains
unclear. As an endocrine organ, adipose tissue secretes multiple bioactive factors, such as leptin tumor necrosis factor-α and interleukin-6, which influence insulin resistance and β-cell dysfunction 6. Adipose tissue is classified as white adipose tissue (WAT) and brown adipose tissue (BAT); in humans, BAT exists only in infants 7. Although both WAT and BAT are related to the development of metabolic disorders, visceral adipose tissue (VAT), a subtype of WAT, is considered more metabolically active, more insulin resistant and a greater contributor to metabolic disorders than BAT
8-11
.
Therefore, studying VAT will be helpful to elucidate the causes of insulin resistance. Proteomics has been widely used in diabetes studies, and proteomic approaches have been applied to analyze adipose tissue
12-14
. Proteomic studies on T2DM-related
adipose tissues have been performed, resulting in identifications of differentially expressed proteins involved in cytoskeleton function and structure, oxidative stress, inflammation and retinoid metabolism15-19. - However, only a few proteomic studies have focused on GDM. Oliva et al 22 identified 14 differentially expressed proteins in 3
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omental adipose tissue from women with GDM compared to women with normal glucose tolerance (NGT) using 2D differential in-gel electrophoresis (2D-DIGE). As a gel-based technique, 2D-DIGE has many limitations, such as lack of automation, poor loading capacity, limited detection capability for hydrophobic and low-abundance proteins, and poor separation of acidic and basic proteins
23, 24
. Therefore, to better
understand the mechanisms of insulin resistance in GDM adipose tissue, a label-free quantitative proteomic approach was employed in this study. Label-free quantitative proteomics is a gel-free method applied to quantitatively analyze protein samples without any isotopic labeling 25. According to data from the Association of Biomolecular Resource Facilities Proteomics Research Group, the observed ratios of proteins quantified using label-free quantitative proteomics are close to the expected values, especially for low-abundance proteins in mixed samples 26, 27
. Therefore, the label-free quantitative proteomic approach, as a useful tool to
measure changes in protein abundance between different samples, has been widely used in clinical studies 28. In this study, a total of 3528 proteins were identified in omental adipose tissue samples from GDM patients and NGT controls using label-free quantitative proteomics. Of these, 66 proteins were significantly different (p-value
