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The Role of the BH4 Cofactor in Nitric Oxide Synthase Activity and Cancer Progression: Two Sides of the Same Coin

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The Role of the BH4 Cofactor in Nitric Oxide Synthase Activity and Cancer Progression: Two Sides of the Same Coin International Journal of Molecular Sciences Review The Role of the BH4 Cofactor in Nitric Oxide Synthase Activity and Cancer Progression: Two Sides of the Same Coin Diego Assis Gonçalves 1,2 , Miriam Galvonas Jasiulionis 3 and Fabiana Henriques Machado de Melo 4,5,* 1 Micro-Imuno-Parasitology Department, Universidade Federal de São Paulo, São Paulo 04023-062, Brazil; [email protected] 2 Department of Parasitology, Microbiology and Immunology, Federal University of Juiz de Fora, Juiz de Fora 36036-900, Brazil 3 Pharmacology Department, Universidade Federal de São Paulo, São Paulo 04023-062, Brazil; [email protected] 4 Department of Pharmacology, Institute of Biomedical Science, University of São Paulo, São Paulo 05508-000, Brazil 5 Institute of Medical Assistance to Public Servants of the State (IAMSPE), São Paulo 04039-000, Brazil * Correspondence: [email protected] or [email protected] Abstract: Cancer development is associated with abnormal proliferation, genetic instability, cell death resistance, metabolic reprogramming, immunity evasion, and metastasis. These alterations are triggered by genetic and epigenetic alterations in genes that control cell homeostasis. Increased reactive oxygen and nitrogen species (ROS, RNS) induced by different enzymes and reactions with distinct molecules contribute to malignant transformation and tumor progression by modifying DNA, proteins, and lipids, altering their activities. Nitric oxide synthase plays a central role in oncogenic signaling modulation and redox landscape. Overexpression of the three NOS isoforms has been Citation: Gonçalves, D.A.; found in innumerous types of cancer contributing to tumor growth and development. Although Jasiulionis, M.G.; Melo, F.H.M.d. The the main function of NOS is the production of nitric oxide (NO), it can be a source of ROS in some Role of the BH4 Cofactor in Nitric pathological conditions. Decreased tetrahydrobiopterin (BH4) cofactor availability is involved in Oxide Synthase Activity and Cancer NOS dysfunction, leading to ROS production and reduced levels of NO. The regulation of NOSs Progression: Two Sides of the Same by BH4 in cancer is controversial since BH4 has been reported as a pro-tumoral or an antitumoral Coin. Int. J. Mol. Sci. 2021, 22, 9546. molecule. Therefore, in this review, the role of BH4 in the control of NOS activity and its involvement https://doi.org/10.3390/ in the capabilities acquired along tumor progression of different cancers was described. ijms22179546 Keywords: Academic Editor: Tzong-Shyuan Lee cancer; nitric oxide synthase; tetrahydrobiopterin; sepiapterin Received: 1 August 2021 Accepted: 23 August 2021 Published: 2 September 2021 1. Background Cancers are a set of diseases characterized by genetic instability, abnormal cell prolifer- Publisher’s Note: MDPI stays neutral ation, cell death resistance, metabolic reprogramming, angiogenesis, metastasis capability, with regard to jurisdictional claims in and immune response evasion caused by genetic and epigenetic alterations in oncogenes published maps and institutional affil- and tumor suppressor genes [1]. These alterations contribute to malignant transformation iations. and tumor development with the consequent acquisition of an increasingly aggressive phenotype through dysregulation of signaling pathways that maintain cell homeostasis. Despite the advancement of medical technology and the development of new target ther- apies [2,3], the incidence of different cancers is rising worldwide and will continue to Copyright: © 2021 by the authors. increase over the course of this century according to the International Agency for the Licensee MDPI, Basel, Switzerland. Research on Cancer (IARC) GLOBOCAN database within the Global Cancer Observa- This article is an open access article tory (COSMIC v94, released 28-MAY-21). This scenario is mainly attributed to high life distributed under the terms and expectancy, the obesity epidemic, increased ultraviolet (UV) radiation exposure, and infec- conditions of the Creative Commons tious pathogens classified as human carcinogens [4–6]. Attribution (CC BY) license (https:// Among other cause effects, tobacco, obesity, and related metabolic syndromes, infec- creativecommons.org/licenses/by/ tions, and UV are associated with inflammation, loss of redox homeostasis, and oxidative 4.0/). Int. J. Mol. Sci. 2021, 22, 9546. https://doi.org/10.3390/ijms22179546 https://www.mdpi.com/journal/ijms Int. J. Mol. Sci. 2021, 22, x FOR PEER REVIEW 2 of 22 Among other cause effects,tobacco, obesity, and related metabolic syndromes, infec- tions, and UV are associated with inflammation, loss of redox homeostasis, and oxidative stress, contributing to cancer development [7–10]. A pro-oxidant milieu alters redox sig- Int. J. Mol. Sci. 2021, 22, 9546 2 of 21 naling pathways improving the acquisition of cancer-related hallmarks. Moreover, nitric oxide (NO) and reactive nitrogen species (RNS) metabolism are also altered in inflamma- tion [11]. High reactive oxygen species (ROS) levels found in tumors are caused by differ- stress,ent mechanisms, contributing including to cancer increased development NADPH [7– 10oxidase]. A pro-oxidant expression milieuand activity, alters mitochon- redox sig- nalingdrial damage, pathways endoplasmic improving reticulum the acquisition stress, of and cancer-related nitric oxide synthase hallmarks. dysfunction Moreover, (NOS) nitric oxide[12–16] (NO). and reactive nitrogen species (RNS) metabolism are also altered in inflam- mation [11]. High reactive oxygen species (ROS) levels found in tumors are caused by different2. Nitric Oxide mechanisms, Synthase including increased NADPH oxidase expression and activity, mito- chondrialThree damage, NOS (EC endoplasmic 1.14.13.39) isoforms reticulum have stress, been and described nitric oxide in mammalian synthase dysfunction cells. NOS (NOS) [12–16]. isoforms, named neuronal NOS (NOS1), endothelial NOS (NOS3), and inducible NOS 2.(NOS2 Nitric), Oxideare encoded Synthase by different genes localized in distinct chromosomes. All three isoforms are homodimeric and each monomer contains a C-terminal reductase domain Three NOS (EC 1.14.13.39) isoforms have been described in mammalian cells. NOS iso- and an N-terminal oxygenase domain connected by a calmodulin-binding peptide linker. forms, named neuronal NOS (NOS1), endothelial NOS (NOS3), and inducible NOS (NOS2), The binding sites for the cofactors flavin mononucleotide (FMN), flavin adenine dinucle- are encoded by different genes localized in distinct chromosomes. All three isoforms are ho- otide (FAD), and nicotinamide adenine dinucleotide phosphate (NADPH) are found in modimeric and each monomer contains a C-terminal reductase domain and an N-terminal the reductase domain, which shares homology with cytochrome P450 (CYT P450) reduc- oxygenase domain connected by a calmodulin-binding peptide linker. The binding sites for tases while the oxygenase domain contains binding sites for heme, the cofactor (6R)- the cofactors flavin mononucleotide (FMN), flavin adenine dinucleotide (FAD), and nicoti- 5,6,7,8-tetrahydrobiopterin (BH4) and the substrate L-arginine. The electron transfer oc- namide adenine dinucleotide phosphate (NADPH) are found in the reductase domain, curs from the NADPH cofactor into the oxygenase domain, where the L-arginine hydrox- which shares homology with cytochrome P450 (CYT P450) reductases while the oxyge- ylation and subsequently NO formation takes place. Stable dimerization between the two nase domain contains binding sites for heme, the cofactor (6R)-5,6,7,8-tetrahydrobiopterin (BH4)NOS monomers and the substrate is required L-arginine. for efficient The electronNOS activity, transfer as occurselectrons from flow the from NADPH the reductase cofactor intodomain the of oxygenase one monomer domain, to the where oxidase the domain L-arginine localized hydroxylation in the other and monomer subsequently (Figure NO 1) formation[17,18]. NOSs takes activity place. Stableis regulated dimerization by integrating between mechanisms, the two NOS including monomers p isost required-transla- fortional efficient modifications, NOS activity, protein as– electronsprotein interactions, flow from the and reductase substrate domain and cofactor of one availability, monomer tobeing the oxidaseBH4 concentration domain localized determinant in the [19 other–21] monomer. (Figure1)[ 17,18]. NOSs activity is regulated by integrating mechanisms, including post-translational modifications, protein– protein interactions, and substrate and cofactor availability, being BH4 concentration determinant [19–21]. Figure 1. Nitric oxide synthase (NOS) activity. NOSs are homodimers and each monomer contains a reductase domain and an oxygenase domain connected by a calmodulin-binding peptide linker (CaM) (for eNOS and nNOS). In the normal NOS coupled state, electron (e−) flow through the nicotinamide adenine dinucleotide phosphate (NADPH) and flavin domains, adenineFigure 1. dinucleotide Nitric oxide (FAD), synthase and (NOS) flavin mononucleotideactivity. NOSs are (FMN), homodimers from the and reductase each monomer domain of contains one monomer a reductase to the domain iron of theand heme an oxygenase group (Fe) domain localized connected in oxidase by domaina calmodulin of the-binding other monomer. peptide linker (6R)-5,6,7,8-tetrahydrobiopterin (CaM) (for eNOS and nNOS). (BH4) In the binding normal to theNOS heme coupled active state, site at electron the interface (e−) flow between through the two the monomersnicotinamide increased
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