Gas chromatography/mass spectrometric assay of endogenous cellular lipid peroxidation products: quantitative analysis of 9‐ and 10‐hydroxystearic acids

Bertucci, Carlo; Hudaib, Mohammad; Boga, Carla; Calonghi, Natalia; Cappadone, Concettina; Masotti, Lanfranco

doi:10.1002/rcm.656

Cited by 15 publications

(12 citation statements)

References 11 publications

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“…Synthesis of 9-HSA and 9-deutero-9-hydroxystearic acid Methyl 9-oxooctadecanoate and methyl 9-hydroxyoctadecanoate were prepared according to the literature (12). 1 H-and 13 C-NMR spectra were recorded on a Gemini 300 spectrometer (Varian, Palo Alto, CA) at 300 and 75.46 MHz, respectively.…”

Section: Methodsmentioning

confidence: 99%

Histone deacetylase 1: a target of 9-hydroxystearic acid in the inhibition of cell growth in human colon cancer

Calonghi

Cappadone

Pagnotta

et al. 2005

Journal of Lipid Research

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Recent studies have shown that an endogenous lipoperoxidation product, 9-hydroxystearic acid (9-HSA), acts in colon carcinoma cells (HT29) as a growth inhibitor by inducing p21 WAF1 in an immediate-early, p53-independent manner and that p21 WAF1 is required for 9-HSA-mediated growth arrest in HT29 cells. It is conceivable, therefore, to hypothesize that the cytostatic effect induced by this agent is at least partially associated with a molecular mechanism that involves histone deacetylase 1 (HDAC1) inhibition, as demonstrated for sodium butyrate and other specific inhibitors, such as trichostatin A and hydroxamic acids. Here, we show that, after administration, 9-HSA causes an accumulation of hyperacetylated histones and strongly inhibits the activity of HDAC1. The interaction of 9-HSA with the catalytic site of the enzyme has been highlighted by computational modeling of the human HDAC1, using its homolog from the hyperthermophilic Aquifex aeolicus as a template. Consistent with the experimental data, we find that 9-HSA can bind to the active site of the protein, showing that the inhibition of the enzyme can be explained at the molecular level by the ligand-protein interaction. Supplementary key words endogenous lipid peroxidation products • tumor • mass spectrometry • computational modeling 9-Hydroxystearic acid (9-HSA) belongs to the class of endogenous lipid peroxidation by-products that are greatly diminished in tumors and therefore become unable to exert their normal controlling functions on cell division (1). Indeed, in HT29 cells, exogenous administration of 9-HSA at micromolar concentrations results in a significant inhibition of proliferation rate as well as a significant increase of p53-independent p21 WAF1 expression (2). The expression of the cell cycle kinase inhibitor p21 WAF1 is induced in neoplastic cells by histone deacetylase 1 (HDAC1) inhibitors such as phenyl butyrate, sodium butyrate, trichostatin A (TSA), and suberoylanilide hydroxamic acid (SAHA). Increased expression of p21 WAF1 may play a critical role in the growth arrest induced in transformed cells by these agents, and it can be regulated, at least in part, by histone acetylation of the chromatin associated with the p21 WAF1 gene. Histone acetylation and gene activation induced by HDAC1 inhibitors would consequently be selective (3). The mode of action of 9-HSA is similar to that of HDAC inhibitors.Crystallographic studies performed using TSA and SAHA indicate that these compounds inhibit HDAC activity by interacting with the catalytic site, thereby blocking substrate access (4, 5). Short-chain fatty acids, such as phenyl butyrate and phenyl acetate, inhibit HDAC activity and affect the expression of numerous genes with disparate cellular functions (6)(7)(8). These agents have been tested in the clinic, but they suffer from a short plasma half-life as well as from the relatively high (millimolar) concentrations that are required for their action. On the other hand, hydroxamic acids such as TSA, SAHA, m -carboxycinnamic acid ...

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Section: Methodsmentioning

confidence: 99%

Histone deacetylase 1: a target of 9-hydroxystearic acid in the inhibition of cell growth in human colon cancer

Calonghi

Cappadone

Pagnotta

et al. 2005

Journal of Lipid Research

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“…A downstream product of ROS is the peroxidized lipid 9-hydroxystearic acid (9-HSA), which we found endogenously produced in human embryonic intestine cells and colon adenocarcinoma as a signaling messenger produced by lipid peroxidation. The amounts of 9-HSA in cancer cells inversely correlating with tumor growth, making it a candidate endogenous mediator of redox signaling in proliferative tissues (Bertucci et al, 2002;Calonghi et al, 2007;Cavalli et al, 1991;Gesmundo et al, 1994;Piretti et al, 1987;Piretti and Pagliuca, 1989).…”

Section: Introductionmentioning

confidence: 99%

Redox Signaling via Lipid Peroxidation Regulates Retinal Progenitor Cell Differentiation

et al. 2019

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“…To unravel the role of ROS scavenging and lipid peroxidation in RPCs, we investigated the in vivo function of 9 -HSA, a known downstream product of lipid peroxidation that we previously demonstrated as a regulator of cellular growth in different human tumor cell lines (Bertucci et al , 2002; Calonghi et al , 2007; Cavalli et al , 1991; Gesmundo et al , 1994). In these studies, we revealed by biochemical and structural modeling studies that, in vitro , 9 -HSA inhibits histone deacetylase 1 (HDAC1) activity by directly binding to its catalytic site (Calonghi et al , 2005; Parolin et al , 2012).…”

Section: Discussionmentioning

confidence: 99%

“…We previously characterized the in vitro role of the 9 -hydroxystearic acid ( 9 -HSA), which was found endogenously produced in human embryonic intestine cells and colon adenocarcinoma as signaling messenger produced by lipid peroxidation (Piretti et al , 1987; Piretti & Pagliuca, 1989). 9 -HSA amounts in cancer cells were found inversely correlating with tumor growth (Bertucci et al , 2002; Calonghi et al , 2007; Cavalli et al , 1991; Gesmundo et al , 1994). We found that in zebrafish, 9 -HSA is endogenously produced and its exogenous administration to RPCs severely altered their differentiation, identifying it as a critical molecule acting downstream of redox signaling events.…”

Section: Introductionmentioning

confidence: 99%

Redox signalingvialipid peroxidation regulates retinal progenitor cell differentiation

Albadri

Naso

Gauron

et al. 2019

Preprint

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21Reactive oxygen species (ROS) and downstream products of lipid oxidation are emerging as important 22 secondary messengers in tissue homeostasis. However their regulation and mechanism of action remain 23 poorly studied in vivo during normal development. Here we reveal that the fine regulation of hydrogen 24 peroxide (H2O2) levels at the degradation step by its scavenger Catalase is crucial to mediate the switch 25 from proliferation to differentiation in retinal progenitor cells (RPCs). We further show that altering the 26 levels of downstream products of the Redox signaling can also affect this switch. Indeed, we identify 9-27 hydroxystearic acid (9-HSA), an endogenous downstream lipid peroxidation product, as a mediator of 28 this effect in the zebrafish retina. In fact, RPCs exposed to higher amounts of 9-HSA failed to 29 differentiate and remained proliferative. We found that 9-HSA exerts its biological function in vivo by 30 inhibiting the activity of histone deacetylase 1. We finally show that the local and temporal 31 manipulation of H2O2 levels by catalase overexpression in RPCs was sufficient to trigger their premature 32 differentiation. Therefore the amount of H2O2 in RPCs is instructive of their ability to switch from 33 proliferation to differentiation. We propose a mechanism that acts in RPC and linking H2O2 homeostasis 34 and neuronal differentiation via the modulation of lipid peroxidation. 35 36 37 38 39 40 41 42 43 44 45 46 Key words 47 48 Redox signaling, lipid peroxidation, retinal progenitor cells, stem cell metabolism, neuronal 49 differentiation, H2O2, 9-HSA, HDAC1, zebrafish 50 4 retinogenesis, retinal progenitor cells (RPCs) start differentiating in a central to peripheral manner. By 3 81 dpf, the central retina is fully differentiated and a peripheral stem cell niche, known as the ciliary 82 marginal zone (CMZ), forms to ensure tissue homeostasis throughout the lifelong growth of the fish. The 83 CMZ comprises three main cell domains labeled by the expression of different transcription factors: the 84 rx2 domain, which contains RSCs (retinal stem cells) and RPCs, the ccnd1 (zebrafish equivalent to 85 cyclinD1) domain where cycling progenitors are found and an ath5 domain, which contains cycling and 86 committed RPCs (Agathocleous & Harris, 2009; Ohnuma & Harris, 2003; Reinhardt et al, 2015; Wan et 87 al, 2016). Due to this precise spatial and temporal organization, the zebrafish retina provides an excellent 88 model to assess the effects of ROS and lipid peroxidation downstream products on a stem cell and 89 progenitor cell population in vivo. Here we show that H2O2 levels are dynamically regulated in the 90 developing and in the mature neuro-retina where the expression of the H2O2 scavenger catalase follows a 91 pattern reminiscent of the neurogenic progression. H2O2 levels are further reflected on the lipid 92 peroxidation state in these cells as shown by the enrichment in the end marker of lipid peroxidation 4-93 hydroxynonenal (4-HNE) within the CMZ. Thus, H2O2 and lipid peroxidat...

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Gas chromatography/mass spectrometric assay of endogenous cellular lipid peroxidation products: quantitative analysis of 9‐ and 10‐hydroxystearic acids

Cited by 15 publications

References 11 publications

Histone deacetylase 1: a target of 9-hydroxystearic acid in the inhibition of cell growth in human colon cancer

Histone deacetylase 1: a target of 9-hydroxystearic acid in the inhibition of cell growth in human colon cancer

Redox Signaling via Lipid Peroxidation Regulates Retinal Progenitor Cell Differentiation

Redox signalingvialipid peroxidation regulates retinal progenitor cell differentiation

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