Transfection and Functional Expression of CYP4A1 and CYP4A2 Using Bicistronic Vectors in Vascular Cells and Tissues

Wang, Ji Shi; Zhang, Fan; Jiang, Miao; Wang, Mong Heng; Zand, Barbara A.; Abraham, Nader G.; Nasjletti, Alberto; Laniado−Schwartzman, Michal

doi:10.1124/jpet.104.070979

Cited by 11 publications

(6 citation statements)

References 25 publications

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“…Cells were infected with the adenoviruses; GFP and CYP4A2 protein levels were measured by Western blot and CYP catalytic activity was evaluated by either measuring -hydroxylation of the CYP4A-preferred substrate, lauric acid, 24 or by quantifying endogenous 20-HETE levels. 25 …”

Section: In Vitro Transductionmentioning

confidence: 99%

Endothelial Dysfunction and Hypertension in Rats Transduced With CYP4A2 Adenovirus

Wang

Singh

Zhang

et al. 2006

Circulation Research

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112

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Abstract-Vascular cytochrome P450 (CYP) 4A enzymes catalyze the synthesis of 20-hydroxyeicosatetraenoic acid (20-HETE), an eicosanoid which participates in the regulation of vascular tone by sensitizing the smooth muscle cells to constrictor and myogenic stimuli. This study was undertaken to investigate the consequences of CYP4A overexpression on blood pressure and endothelial function in rats treated with adenoviral vectors carrying the CYP4A2 construct. Intravenous injection of Adv-CYP4A2 increased blood pressure (from 114Ϯ1 to 133Ϯ1 mm Hg, PϽ0.001), and interlobar renal arteries from these rats displayed decreased relaxing responsiveness to acetylcholine, which was offset by treatment with an inhibitor of CYP4A. Relative to data in control rats, arteries from Adv-CYP4A2-transduced rats produced more 20-HETE (129Ϯ10 versus 97Ϯ7 pmol/mg protein, PϽ0.01) and less nitric oxide (NO; 4.2Ϯ1.6 versus 8.4Ϯ1 nmol nitriteϩnitrate/mg; PϽ0.05). They also displayed higher levels of oxidative stress as measured by increased generation of superoxide anion and increased expression of nitrotyrosine and gp91phox. Collectively, these findings demonstrate that augmentation in vascular 20-HETE promotes the development of hypertension and causes endothelial dysfunction, a condition characterized by decreased NO synthesis and/or bioavailability, imbalance in the relative contribution of endothelium-derived relaxing and contracting factors, and enhanced endothelial activation. T he endothelium plays a critical role in the short-and long-term regulation of the cardiovascular system. It serves as a protective barrier between tissues and circulating blood and functions to maintain vascular homeostasis by releasing bioactive factors in response to hemodynamic changes and blood borne signals. Such endothelial cell functions are impaired in many disease processes, including, diabetes, atherosclerosis, and hypertension.The expression of the cytochrome P450 (CYP) 4A and its catalytic activity as arachidonic acid -hydroxylase which produces 20-hydroxyeicosatetraenoic acid (20-HETE) has been linked to the regulation of vascular reactivity and tone and to the development of hypertension. This notion is substantiated by observations that 20-HETE promotes vasoconstriction, 1,2 that CYP4A expression and/or 20-HETE production is increased in vascular tissues of hypertensive animals, 3 and that interventions that decrease CYP4A expression and/or activity cause blood pressure to fall in animal models of hypertension. 4 -11 The vasoconstrictor action of 20-HETE is primarily attributed to inhibition of the large conductance Ca 2ϩ -activated K Their study demonstrated that 20-HETE attenuates acetylcholine-induced dilation in cremasteric arterioles of normotensive rats on a low salt diet and that inhibition of 20-HETE synthesis enhances arteriolar dilation to acetylcholine in hypertensive rats on a high salt diet. More recently, 20-HETE has been shown to functionally antagonize EDHF-mediated relaxation in small porcine coronary arteries. 20 In the pre...

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Section: In Vitro Transductionmentioning

confidence: 99%

Endothelial Dysfunction and Hypertension in Rats Transduced With CYP4A2 Adenovirus

Wang

Singh

Zhang

et al. 2006

Circulation Research

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112

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“…In mammalian cells, the most studied and effective subfamily to produce 20-HETE is CYP4A [ 53 ]. In rats, there are four isoforms identified: CYP4A1, CYP4A2, CYP4A3, and CYP4A8 [ 89 ]. These isoforms share 66–98% homology and common catalytic activity and are expressed in the liver, kidney, and brain [ 90 ].…”

Section: Arachidonic Acid Metabolismmentioning

confidence: 99%

Arachidonic Acid Metabolite as a Novel Therapeutic Target in Breast Cancer Metastasis

Borin

Angara

Rashid

et al. 2017

IJMS

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Metastatic breast cancer (BC) (also referred to as stage IV) spreads beyond the breast to the bones, lungs, liver, or brain and is a major contributor to the deaths of cancer patients. Interestingly, metastasis is a result of stroma-coordinated hallmarks such as invasion and migration of the tumor cells from the primary niche, regrowth of the invading tumor cells in the distant organs, proliferation, vascularization, and immune suppression. Targeted therapies, when used as monotherapies or combination therapies, have shown limited success in decreasing the established metastatic growth and improving survival. Thus, novel therapeutic targets are warranted to improve the metastasis outcomes. We have been actively investigating the cytochrome P450 4 (CYP4) family of enzymes that can biosynthesize 20-hydroxyeicosatetraenoic acid (20-HETE), an important signaling eicosanoid involved in the regulation of vascular tone and angiogenesis. We have shown that 20-HETE can activate several intracellular protein kinases, pro-inflammatory mediators, and chemokines in cancer. This review article is focused on understanding the role of the arachidonic acid metabolic pathway in BC metastasis with an emphasis on 20-HETE as a novel therapeutic target to decrease BC metastasis. We have discussed all the significant investigational mechanisms and put forward studies showing how 20-HETE can promote angiogenesis and metastasis, and how its inhibition could affect the metastatic niches. Potential adjuvant therapies targeting the tumor microenvironment showing anti-tumor properties against BC and its lung metastasis are discussed at the end. This review will highlight the importance of exploring tumor-inherent and stromal-inherent metabolic pathways in the development of novel therapeutics for treating BC metastasis.

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“…20-HETE, the ω-hydroxylation product of AA, is the principal AA metabolite of CYP enzymes in vascular smooth muscle [21] and kidney [22]. Synthesis of 20-HETE is catalyzed by the CYP4A gene family [21] (Figure 2).…”

Section: Regulation Of Cerebral Vascular Function By Cox- and Cyp-mentioning

confidence: 99%

“…Synthesis of 20-HETE is catalyzed by the CYP4A gene family [21] (Figure 2). This subfamily encodes several CYP enzymes in different species.…”

Section: Regulation Of Cerebral Vascular Function By Cox- and Cyp-mentioning

confidence: 99%

Cyclooxygenase- and cytochrome P450-derived eicosanoids in stroke

Huang

Al‐Shabrawey

Wang

2016

Prostaglandins & Other Lipid Mediators

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Arachidonic acid (AA) is metabolized by cyclooxygenase (COX) and cytochrome P450 (CYP) enzymes into eicosanoids, which are involved in cardiovascular diseases and stroke. Evidence has demonstrated the important functions of these eicosanoids in regulating cerebral vascular tone, cerebral blood flow, and autoregulation of cerebral circulation. Although COX-2 inhibitors have been suggested as potential treatments for stroke, adverse events, including an increased risk of stroke, occur following long-term use of coxibs. It is important to note that prolonged treatment with rofecoxib increased circulating levels of 20-hydroxyeicosatetraenoic acid (20-HETE), and 20-HETE blockade is a possible strategy to prevent coxib-induced stroke events. It appears that 20-HETE has detrimental effects in the brain, and that its blockade exerts cerebroprotection against ischemic stroke and subarachnoid hemorrhage (SAH). There is clear evidence that activation of EP2 and EP4 receptors exerts cerebroprotection against ischemic stroke. Several elegant studies have contributed to defining the importance of stabilizing the levels of epoxyeicosatrienoic acids (EETs), by inhibiting or deleting soluble epoxide hydrolase (sEH), in stroke research. These reports support the notion that sEH blockade is cerebroprotective against ischemic stroke and SAH. Here, we summarize recent findings implicating these eicosanoid pathways in cerebral vascular function and stroke. We also discuss the development of animal models with targeted gene deletion and specific enzymatic inhibitors in each pathway to identify potential targets for the treatment of ischemic stroke and SAH.

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Transfection and Functional Expression of CYP4A1 and CYP4A2 Using Bicistronic Vectors in Vascular Cells and Tissues

Cited by 11 publications

References 25 publications

Endothelial Dysfunction and Hypertension in Rats Transduced With CYP4A2 Adenovirus

Endothelial Dysfunction and Hypertension in Rats Transduced With CYP4A2 Adenovirus

Arachidonic Acid Metabolite as a Novel Therapeutic Target in Breast Cancer Metastasis

Cyclooxygenase- and cytochrome P450-derived eicosanoids in stroke

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