@article{ead4a17489644bd3adbb5e6c2d38992b,
title = "Sex-specific extracellular matrix remodeling during early adipogenic differentiation by gestational bisphenol A exposure",
abstract = "Bisphenol A (BPA) is an endocrine disrupting chemical known to promote adipose tissue mass in vivo and adipogenesis in vitro. Whether BPA can affect and reprogram early adipogenic differentiation signals that trigger adipogenic differentiation, remains unknown. We hypothesized that gestational BPA exposure results in a preadipocyte phenotype that leads to accelerated adipogenic differentiation, and that this phenotype is sex specific. Primary ovine fetal preadipocytes were derived from control (C) and BPA-exposed during pregnancy and differentiated in vitro. Gestational BPA enhanced lipid accumulation at early stages of differentiation (48 h) and this was evident in females but not male-derived fetal preadipocytes. After an RNA sequencing approach, samples were compared as follows: 2 groups (C vs. BPA); 2 sexes (female (F) vs. male (M)); and 2 time points (0 h vs. 48 h). Before differentiation, 15 genes were differentially expressed between the C and the BPA-exposed preadipocytes within sex. In BPA-F, extracellular matrix remodeling genes cathepsin K and collagen 5α3 were upregulated compared to C–F. At 48 h, BPA-F had 154 genes differentially expressed vs. C–F and BPA-M had 487 genes differentially expressed vs. C-M. Triglyceride and glycerophospholipid metabolism were the most upregulated pathways in BPA-F. Downregulated pathways were associated with extracellular matrix organization in BPA-exposed preadipocytes. These findings are among the first to demonstrate that gestational BPA can modify the fate of adipocyte precursors by altering pathways associated to extracellular matrix components, an often-disregarded, but required aspect of adipogenic differentiation. This work highlights the need to investigate early adipogenic differentiation changes in other obesogenic chemicals.",
keywords = "Adipogenesis, Bisphenol A, Fetal, Preadipocyte",
author = "Yong Pu and Elvis Ticiani and Waye, {Anita A.} and Kunzhe Dong and Huanmin Zhang and Almudena Veiga-Lopez",
note = "Funding Information: The transcriptomic analysis revealed that in female-derived preadipocytes, gestational BPA exposure triggered the upregulation of extracellular matrix proteins involved in preadipocyte differentiation, such as CTSK, Col5α3, and CYGB. Known to have a collagenolytic activity capable of degrading collagen I (Chiellini et al., 2003), CTSK is upregulated in white adipose tissue of rodent models of obesity and obese humans (Chiellini et al., 2003; Funicello et al., 2007). The observation that CTSK-null mice in the postweaning period and CTSK-null mice exposed to a high-fat diet show reduced adiposity (Funicello et al., 2007), suggests that CTSK collagenolytic activity plays a role in white adipose tissue development. Therefore, BPA-induced CTSK upregulation in undifferentiated preadipocytes supports CTSK's adipogenic role during differentiation. Because CTSK can also be modulated by BPA in other organ systems (Chiellini et al., 2003), further studies should be conducted to demonstrate if BPA can directly modulate CTSK's transcription. While BPA exerts affinity for the estrogen receptor (ESR1) (Hiroi et al., 1999; Cao et al., 2017), it is unlikely that its effect on CTSK expression is ESR1-mediated, since estradiol has been reported to downregulate CTSK expression (Mano et al., 1996; Troen, 2006). Most of collagens are degraded by CTSK during preadipocyte differentiation (Mano et al., 1996). However, Col5α3, which is highly expressed in white adipose tissue (Huang et al., 2011), is upregulated upon adipogenic differentiation (Huang et al., 2011). Also, Col5a3 chains, do not appear to increase the rigidity of extracellular matrix collagenous meshwork, thus allowing a better microenvironment for the preadipocyte differentiation (Khan et al., 2009), even in cell culture conditions (Kubo et al., 2000). Thus, the higher expression of Col5α3 may support the higher lipid accumulation observed in BPA exposed female preadipocytes.CYGB, a globin family member protein, is expressed in several soft tissues, including white adipose tissue (Ayyappan and Nagajyothi, 2017). With a known role in the production of extracellular proteins such as collagen (Nakatani et al., 2004; Tateaki et al., 2004), CYGB has also been reported to be upregulated in late-stages of adipogenic differentiation with a parallel profile to that of classic adipogenic differentiation genes, such as CEBPa and FABP4 (Dogan et al., 2017). Interestingly, with a tendency in the qPCR analyses and upregulation in our transcriptomic analyses, changes in CYGB were detectable much earlier in the differentiation process (Day 2) compared to later (Day 8 of differentiation) in previous studies (Dogan et al., 2017). Given that CYGB's expression profile parallels that of FABP4 and that CYGB's overexpression enhances lipid accumulation in 3T3-L1 preadipocytes (Dogan et al., 2017), it is possible that CYGB's upregulation in BPA-exposed female preadipocytes contributed to their enhanced adipogenic differentiation. Given that BPA can induce methylation changes on the PPARγ promoter (Longo et al., 2020; Rahmani et al., 2020; Kubota et al., 2021), future studies should investigate if the transcriptomic signature observed in extracellular matrix-related genes are the result of epigenetic modifications in preadipocytes. Altogether, upregulation of genes involved in extracellular matrix remodeling upon exposure to BPA suggests a direct programing effect during fetal life. To note, these findings were restricted to female preadipocytes, in support of a sex specific effect in the adipose tissue as previously described (Somm et al., 2009; Pu et al., 2017; Rubin et al., 2017). On the other hand, estradiol can affect white adipose tissue accumulation (D'Eon et al., 2005; Pallottini et al., 2008; Ishikawa et al., 2020). This, coupled with the fact that BPA activates ESRs (Pelch et al., 2019) and that female fetal preadipocytes have a higher baseline expression of ESR1 and ESR2 (Pu et al., 2017) support the hypothesis that differential expression of ESR between female and male preadipocytes contributes to the sex-specific differences observed in response to BPA exposure.Most of the differential expression was observed during early adipogenic differentiation (Day 2) with over 150 genes being differentially regulated upon gestational BPA exposure. In female preadipocytes, many of the down-regulated genes clustered in pathways related to extracellular matrix organization, including collagens (COL27A1, COL4A4 and COL4A6), integrins (ITGA2, ITGA8, ITGA9 and ITGA11), metallopeptidases (MMP13 and ADAMTS9) and metallopeptidases inhibitors (TIMP1). These observations support previous findings, where the loss of the preadipocyte fibroblastic phenotype is accompanied by 1) a loss in fibrillar collagen types, such as collagen I and III (Weiner et al., 1989) and 2) chemical inhibition of prolyl-hydroxylase, an enzyme required for collagen stability, that impairs triglyceride storage during adipocyte differentiation (Nakajima et al., 2002).Research reported in this publication was supported the National Institute of Environmental Health Sciences of the National Institute of Health (1K22ES026208 and R01ES027863 to A.V-L). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Publisher Copyright: {\textcopyright} 2022 Elsevier Ltd",
year = "2022",
month = sep,
doi = "10.1016/j.chemosphere.2022.134806",
language = "English (US)",
volume = "302",
journal = "Chemosphere",
issn = "0045-6535",
publisher = "Elsevier Limited",
}