Oxidative stress in retinal pigment epithelium degeneration: from pathogenesis to therapeutic targets in dry age-related macular degeneration : Neural Regeneration Research

Secondary Logo

Journal Logo


Oxidative stress in retinal pigment epithelium degeneration: from pathogenesis to therapeutic targets in dry age-related macular degeneration

Maurya, Meenakshi; Bora, Kiran; Blomfield, Alexandra K.; Pavlovich, Madeline C.; Huang, Shuo; Liu, Chi-Hsiu; Chen, Jing*

Author Information
Neural Regeneration Research 18(10):p 2173-2181, October 2023. | DOI: 10.4103/1673-5374.369098
  • Open



Advances in modern healthcare have significantly increased the average life span of humans in the last two centuries. Although ultimately beneficial, this advanced longevity also brings the diseases associated with aging into the limelight, such as cardiovascular diseases, cancer, dementia, and others. In the eye, aging is associated with visual impairment caused by cataracts and also retinal diseases such as age-related macular degeneration (AMD) (Klein et al., 1992; Vingerling et al., 1995; Friedman et al., 2004). As a major cause of blindness in the elderly population (> 65 years old), AMD affects the macula, the central area of the retina responsible for fine color vision. Worldwide, the number of AMD patients is estimated to be ~200 million in 2020, and with the aging population, the number is expected to increase to 288 million by 2040 (Wong et al., 2014).

Development of AMD occurs progressively over decades and based on the symptoms AMD can be classified into three stages: early, intermediate, and late or advanced (Figure 1A and B). Early and intermediate stages show increasing size of drusen, which are lipid-enriched subretinal deposits, and pigmentary changes in retinal pigment epithelium (RPE), a monolayer of phagocytic and nourishing cells beneath the retina. The yellow subretinal deposits known as drusen were first described in 1854 from the German word “druse” meaning “geode”, yet its connection with AMD was not fully recognized until the 20th century (de Jong, 2016). Now considered a key characteristic feature of AMD, drusen are formed due to gradual extracellular accumulation of lipid, protein, and/or cellular debris between RPE and Bruch’s membrane in an age-dependent manner (Dithmar et al., 2001; Hageman et al., 2001). Currently, the size of drusen serves as a classification marker of the clinical progression or worsening of AMD (Ferris et al., 2013). In addition, there are two types of AMD, dry (atrophic) and wet (neovascular). In dry AMD, which accounts for 80–90% of patients, the late advanced stage is marked by the presence of geographic atrophy (GA), characterized by localized sharply delineated atrophic area of RPE and choroidal vessels. On the other hand, wet AMD, a less common neovascular form of late AMD accounting for about 10–15% of AMD patients, features classic choroidal neovascularization (CNV) with associated subretinal fluid and hemorrhages (Table 1). Both GA and CNV are considered key characteristics of the late or advanced stage of AMD for dry and wet forms respectively. While GA in dry AMD results in gradual loss of vision, CNV in wet AMD frequently results in acute and profound vision loss in elderly patients (Bressler, 2002).

Figure 1:
Schematic representation of pathological and cellular changes in RPE-retina complex with AMD development and progression.(A) Pathological features in AMD eyes compared to normal eyes. Genetic and environmental risk factors can lead to cellular oxidative stress that contributes to the accumulation of drusen, a hallmark of AMD, below RPE. (B) Cellular degeneration in AMD. During aging, accumulated oxidative stress with other detrimental factors may trigger biochemical changes in macromolecular alterations, eventuating in RPE dysfunction, thickening of Bruch’s membrane, and drusen formation in the macular region, initiating early/mid-AMD. As AMD progresses, drusen deposition increases and death of RPE cells occurs as observed in geographic atrophy in late dry AMD, with loss of nearby photoreceptors. In neovascular AMD, pathological overgrowth of choroidal vessels may invade into normally avascular sub-retinal space, and cause leakage and bleeding and eventual vision loss. AMD: Age-related macular degeneration; RPE: retinal pigment epithelium. Created with BioRender.com.
Table 1:
Stages of AMD progression and classification

Currently, there is no cure for AMD nor successful ways to prevent its onset. Available treatment options for neovascular AMD include photodynamic therapy, laser photocoagulation, and anti-vascular endothelial growth factor (VEGF) therapies that have been successful in the past two decades, although some patients remain unresponsive to treatment. For the more prevalent dry AMD, ongoing trials include anti-complement, stem cell and RPE replacement therapies; however, few treatments are available to slow down the disease progression, with the only evidence-based means being dietary intake of antioxidant nutrients. The use of antioxidant vitamins is endorsed by promising findings from two Age-Related Eye Disease Study (AREDS) trials, which exhibited their protective effects on reducing the risk of progression to advanced AMD by ~25% over 5 years (AREDS, 2001; AREDS2, 2013). These trial results highlight the important role of countering oxidative stress and damage in dry AMD pathogenesis and disease prevention, although the AREDS formula is not universally recommended by a few other studies with no effects or unanticipated health risks (Evans, 2008; Datta et al., 2017), which potentially reflects the interaction of genetics and vitamin supplement (Vavvas et al., 2018).

AMD is a multifactorial disease with many factors in addition to aging contributing to the disease progression, including environmental agents and genetic predisposition, involving oxidative stress, chronic inflammation, innate immunity (complement pathway), and lipid metabolisms (Jun et al., 2019; Fleckenstein et al., 2021; Zhang and Wong, 2021). The complex interaction among these mechanisms is still under intense investigation to fully elucidate the disease pathogenesis, with the possibility that different pathological processes may take place simultaneously to result in distinct pathologies in multiple AMD-associated cell types (Hadziahmetovic and Malek, 2020; Bird, 2021). This review summarizes the current understanding of AMD pathogenesis with a specific focus on the role of oxidative stress in RPE degeneration in dry AMD. Relevant advances from both clinical and experimental studies are discussed to highlight potential new strategies to counter oxidative damage and protect RPE.

Search Strategy and Selection Criteria

The PubMed database was used to search and select relevant literature, using the following combinations of keywords: age related macular degeneration, antioxidants, oxidative stress, retinal pigment epithelium, Age-Related Eye Disease Study, nuclear factor erythroid-2-related factor 2, and REV-ERBα.

Oxidative Damage Is a Major Cause of Retinal Pigment Epithelium Degeneration in Dry Age-Related Macular Degeneration

RPE plays a vital role in maintaining retinal health

RPE, a monolayer of pigmented epithelial cells between the retina and choroid, has a vital function in supporting photoreceptors and vision with multiple trophic tasks. On the apical side, they recycle photoreceptor outer segments via phagocytosis and assist the visual cycle through retinoid cycling. Apical junctions between RPE cells also form the outer blood-retinal barrier, which selectively transports nutrients and metabolic waste between the neural retina and the choroid, through Bruch’s membrane on the basal side of RPE (Strauss, 2005; Fields et al., 2020). In addition, RPE secretes VEGF towards the choroid to maintain the health of choroidal vessels (Saint-Geniez et al., 2009). RPE also protects the neural retina from light-induced damage through light absorption by melanin and lipofuscin. All these biochemical processes require active oxidative metabolism, hence, making RPE highly susceptible to oxidative damage.

RPE cells are exposed to high levels of reactive oxygen species

Generation of reactive oxygen species (ROS) is an integral part of normal cellular metabolism and physiology, in order to maintain cellular energy demands and function. ROS generation in turn serves as the framework for redox signaling to control its balance. RPE is exposed to high levels of oxidative stress, primarily from excess mitochondrial ROS, and phagocytosis-related oxidative events (Datta et al., 2017; Brown et al., 2018).

The choroid vasculature carries high O2 pressure passing across RPE to the retina, to meet its metabolic demand. RPE contains a large number of mitochondria, which create a highly oxidative environment, with the production of ROS including superoxide, hydrogen peroxide, and hydroxyl radical (Zorov et al., 2014; Brown et al., 2019). In addition to the major ROS-producing site in mitochondrial complexes I & III, complexes II and IV of the electron transport chain system also generate ROS, which leaks from the mitochondrial membrane in normal physiological conditions (Liu et al., 2002). Such ROS leakage tends to increase with aging and can exacerbate the age-dependent ROS accumulation (Cadenas and Davies, 2000; Liang and Godley, 2003; Sohal and Orr, 2012).

Lipid oxidation from phagocytosed outer segments (enriched with polyunsaturated fatty acids), and photo-oxidation due to constant light stimulation, contribute to excess ROS in RPE additionally (Cai et al., 2000; Lambros and Plafker, 2016; Brown et al., 2018). Photoreceptor outer segment disk engulfment is primarily carried out by RPE through a lysosome-efficient system that leads to the accumulation of lipofuscin debris, a marker of cellular senescence, within the lysosome with aging. Environmental and lifestyle factors such as smoking and an unbalanced western diet, serve as additional sources of chemical and dietary oxidants, contributing to high ROS levels in RPE (Malek et al., 2012; Brown et al., 2018; Figure 2).

Figure 2:
Schematic figure illustrating the redox balance between antioxidant pathway and ROS-inducing factors in the generation of oxidative stress causing RPE cell degeneration.To combat ROS and maintain cellular redox homeostasis, RPE cells maintain a powerful NRF2-mediated antioxidant enzyme system, in addition to antioxidant chemicals such as glutathione, as well as Zinc, vitamin-C, -E, Lutein, and zeaxanthin that are part of Age-Related Eye Disease Study supplementation components. The imbalance between ROS generation and antioxidant capacity results in oxidative stress, causing macromolecular (DNA, proteins, and lipid) alterations, thereby initiating intracellular organelle disruption in the form of mitochondrial damage, nuclear degeneration, and eventually RPE dysfunction and death. NRF2: Nuclear factor erythroid-2-related factor 2; ROS: reactive oxidative species; RPE: retinal pigment epithelium. Created with BioRender.com.

RPE redox homeostasis is maintained by an antioxidant self-defense system

To counter the ROS, RPE has well-synchronized signaling mechanisms including antioxidants such as glutathione and heme, antioxidant vitamins, and redox proteins including thioredoxin. Moreover, the elimination of ROS in RPE is carried out primarily by a cascade of nuclear factor erythroid-2-related factor 2 (NRF2)-dependent antioxidant self-defense enzymes, including superoxide dismutase (SOD), catalase, and peroxidase, in addition to free radical scavengers (Cai et al., 2000). This system arms RPE with the ability to quench superoxide and peroxide moieties, along with other free radicals, to maintain cellular homeostasis. During aging this defense mechanism may be impaired or overwhelmed, leading to excess ROS buildup and hence oxidative damage (Suh et al., 2004; Reisman et al., 2009; Gounder et al., 2012; Volonte et al., 2013; Zhang et al., 2015a; Figure 2).

In addition to antioxidants, autophagy may also contribute to the redox defense in RPE cells. In AMD, RPE specifically shows increased ROS levels, susceptibility to lipid peroxide-induced protein modification, and subsequent oxidative stress (Beatty et al., 2000; Kaemmerer et al., 2007). Compared to the normal young retina, autophagosomes, autophagy flux, and autophagy-related proteins such as light chain 3 (LC3)-II/LC3-I ratio levels increase with aging. Gradual accrual of toxic proteins, damaged organelles, and drusen deposits leading to RPE cell dysfunction and death, owing to impaired lysosomal digestion or autophagy has been linked with AMD pathogenesis (Wang et al., 2009; Mitter et al., 2014). In fact, LC3-II/LC3-I ratio fails to increase in late-stage AMD retina and RPE (Golestaneh et al., 2017). Considering the role of autophagy in lipofuscin clearance, diminished autophagic activity might also lead to increased ROS, enhanced lipofuscin accumulation, and subsequent RPE dysfunction and death, hence increasing the risk of GA in AMD.

Oxidative stress might also downregulate RNase DICER1, deficiency of which was suggested to lead to Alu RNA toxicity, NLRP3 inflammasome activation, and RPE atrophy in AMD (Kaneko et al., 2011; Tarallo et al., 2012; Kaarniranta et al., 2020). Given the essential role of DICER1 in microRNA biogenesis, the potential role of miRNA dysregulation in RPE redox control and degeneration is also emerging (Intartaglia et al., 2020; Liu et al., 2020; Urbańska et al., 2022). Several microRNAs show potential in the modulation of RPE homeostasis, function, and autophagy and may become dysregulated in AMD RPE, such as miR-184, miR-29, and miR-1273g (Murad et al., 2014; Cai et al., 2019; Hyttinen et al., 2021) or by their inhibition such as miR-302d-3p (Jiang et al., 2018). Since the present review focuses on the oxidative stress in RPE, readers may refer to recent reviews (Jun et al., 2019; Intartaglia et al., 2020; Hyttinen et al., 2021; Du and Palczewski, 2022; Urbańska et al., 2022) for details on the interplay of microRNAs in oxidative stress and AMD.

Oxidative damage contributes to RPE degeneration

Although reactive oxygen intermediates are mainly the byproducts of physiological processes and part of a normal feedback loop in redox signaling, overexposure of ROS stimuli and impaired antioxidative defense due to aging can result in ROS accumulation. Subsequent oxidative stress and excess free radicals result in oxidative damage to macromolecules such as DNA, proteins, and lipids. In particular, lipid oxidation can alter the normal function of cellular membrane lipid and lipoprotein, and trigger inflammation. Together, these macromolecular oxidative damages can lead to the breakdown of intracellular organelles like mitochondria, and lysosomes, consequently resulting in the death of RPE and photoreceptor cells (Cai et al., 2000; Lambros and Plafker, 2016; Brown et al., 2019; Figure 2).

RPE aging and degeneration lead to vision loss in AMD

Cellular aging manifests as both structural deterioration and compromised function. In RPE, signs of aging can be seen as an alteration in microvilli and basal folding structure, loss of melanin pigment, and accumulation of cellular debris and lipofuscin. These abnormalities in RPE near the macular area that activate the slow RPE dysfunction and degeneration cascade are often considered part of the triggers for AMD development (Bird, 2021).

Since one of the main functions of RPE is the circadian engulfment of the spent rod outer segment (Young and Bok, 1969; LaVail, 1976), any disturbance in this vital recycling process, either due to RPE dysfunction or alienated phagocytosis turnover with aging, may trigger ROS-derived lipid and protein accumulation and cross-link. These molecular changes can also lead to structural and functional changes in mitochondria in RPE. Compared to normal aging RPE, AMD RPE reveals reduced mitochondrial density, reduced adenosine 5’-triphosphate production, and increased loss of cristae with mitochondrial DNA damage (Kaarniranta et al., 2019). In fact, global proteome analysis of RPE from AMD patients confirms the alteration in a vast number of mitochondrial system proteins (Nordgaard et al., 2006), strengthening the idea of RPE mitochondrial dysregulation and damage-driven AMD pathogenesis (Fisher and Ferrington, 2018; Tong et al., 2022).

Bruch’s membrane adjacent to RPE also takes part in this process as age-dependent degeneration in collagen and elastin impairs extracellular matrix maintenance, thereby lowering the rate of extracellular wastage clearance (Spraul et al., 1999). Together with RPE dysfunction, lipid-laden drusen forms, resulting in thickening of Bruch’s membrane (Curcio, 2018; Bird, 2021), which further hinders metabolic exchange between RPE and choroid and may aggravate deposition. Drusen thickening and deposition also trigger chronic inflammation and immune system activation that further worsen the architectural integrity of the RPE-retina complex (Gehrs et al., 2006; Datta et al., 2017), resulting in the eventual loss of trophic support by RPE for adjacent photoreceptors leading to their degeneration or death (Strauss, 2005), and subsequent gradual loss of central vision in AMD (Haddad et al., 2006; Rattner and Nathans, 2006; Figure 1B).

Studies on Environmental and Genetic Risk Factors Support the Pathogenic Role of Oxidative Damage in Age-Related Macular Degeneration

The pathogenic role of oxidative stress is reflected by environmental and lifestyle risk factors of AMD

Numerous environmental factors contribute to accelerated aging and AMD development. The association of sunlight exposure with AMD development was first highlighted by the Beaver Dam Eye Study (Cruickshanks et al., 1993), suggesting the potential risk of photo-oxidation. Apart from aging, smoking acts as a major non-genetic contributing factor in macular atrophy. Smoking is significantly associated with high retinal pigment and exudative macular degeneration in both females (odds ratio [OR]: 2.50) and males (OR: 3.29) active smokers with respect to non-smoker or ex-smoker patients (Klein et al., 1993).

The idea of oxidative factors functioning as an etiological component in AMD development is supported by gene-environmental interaction studies, such as smoking (Smith and Hansch, 2000; Rangasamy et al., 2004). Clinical investigation has suggested that smoking, paired with aging, results in an increased risk of neovascular AMD and vision loss in patients (Chew et al., 2014a). The clinical outcomes were also supported by experimental studies. RPE function requires an immense adenosine 5’-triphosphate supply fulfilled by a large number of mitochondria, with the price of constant exposure in a highly oxidative environment. Chemicals in cigarette smoke cause decreased activity of mitochondrial subunits, and increased mitochondrial superoxide production, resulting in mitochondrial dysfunction (Strunnikova et al., 2004; Jia et al., 2007; Bertram et al., 2009). Chronic exposure to cigarette smoke also compromises the in-built redox machinery within the eye to weaken redox balance. Increased lipid peroxidation (4-HNE) further amplifies oxidative stress and accelerates RPE degeneration (Klein et al., 1998; Cano et al., 2010; Ni Dhubhghaill et al., 2010).

Besides smoking, early AMD development is also accelerated by heavy alcohol consumption and hypertension, all of which are associated with increased oxidative stress and antioxidant imbalance in clinical trials (Seddon et al., 2001; Kuan et al., 2021; Zhang et al., 2021) and are detrimental to RPE. The influence of dietary patterns on AMD was also considered as food is the primary source of dietary vitamins and other antioxidants. The association of early AMD prevalence decreases with plant and seafood-based diet (OR: 0.7) and increases with dairy and refined grain-rich (OR: 1.56), respectively (Chiu et al., 2014). The relative risk (RR) of progression of AMD increases with high-fat consumption, irrespective of source, whether it is plant-based (RR: 2.29) or animal-based (RR: 3.82) high-fat diet (Seddon et al., 2003). The level of lipid saturation also correlates with the likelihood of AMD progression; for saturated fat (RR: 2.09), monounsaturated fat (RR: 2.21), polyunsaturated fat (RR: 2.28), and trans unsaturated fat (RR: 2.39), respectively (Seddon et al., 2003). In fact, smoking (Rahman and MacNee, 1996; Smith and Hansch, 2000; Rangasamy et al., 2004) and high-fat diet intake are considered preventable AMD risk factors (Mares-Perlman et al., 1995; Seddon et al., 2001), and both are linked with increased oxidative stress, supporting the detrimental role of oxidative stress in AMD pathogenesis.

Genetic risk factors of AMD suggest only susceptibility but not causality

Even though aging is the strongest AMD-associated risk factor, many epidemiological studies have highlighted the shared impact of genetics, ethnicity, and family history. The theory of a genetic component of AMD (De Jong et al., 1997; Seddon et al., 1997; Klaver et al., 1998) was supported by studies of monozygotic twins with significantly increased concordance rate compared to dizygotic twins (Dosso and Bovet, 1992; Klein et al., 1994; Meyers, 1994; Hammond et al., 2002; Grizzard et al., 2003). Additionally, AMD prevalence has been associated with ethnicity, i.e., higher in Caucasian populations compared to non-Caucasians (Klein et al., 2006; Bressler et al., 2008). Family history may also play a role, with an OR: 2.4 to 19.8-fold increase in prevalence in individuals with a family history of AMD as compared to those without (Seddon et al., 1997; Smith and Mitchell, 1998), hinting a genetic link.

Advances in molecular genetics provided much-needed evidence since the impact of a genetic component in innate immunity (complement system) on AMD prevalence was discovered and investigated intensely (Zhang and Wong, 2021). Several proteins of the complement system have been identified in drusen found in aging and AMD eye (McHarg et al., 2015). Genes involved in innate immunity, and particularly complement regulation, such as complement factor H (CFH) (Edwards et al., 2005; Haines et al., 2005; Klein et al., 2005), complement 3 (C3), complement 2/complement factor B (Gold et al., 2006; Francis et al., 2009), and complement factor H-related 1, 3 (Spencer et al., 2008) exhibit significant positive association in AMD development and progression. It is estimated that CFH variants, with decreased CFH function, may contribute to approximately 50% of the risk of developing AMD, with much from a missense mutation (Y420H) (Edwards et al., 2005).

In addition to complement, single nucleotide polymorphisms in two closely located, major AMD-associated genes were identified at chromosome 10q26: high-temperature requirement factor A1, serine peptidase 1 (HTRA1) (rs11200638 G→A polymorphism), and age-related maculopathy susceptibility 2 (ARMS2) (rs10490924 G→T polymorphism). Both show strong linkage disequilibrium and increase the risk of AMD by about the same magnitude (Tong et al., 2010), making it difficult to differentiate them. The importance of HTRA1 is supported by its cellular localization within the RPE and drusen in the AMD eyes (Yang et al., 2006). Increased expression of HTRA1, a serine protease, may potentially compromise the structural integrity of Bruch’s membrane, and promote RPE atrophy, whereas ARMS2, a mitochondrial protein, has an unknown function (Kanda et al., 2010). Additional genes with different cellular functions are also associated with AMD risk. For instance, genes related to lipid metabolism pathways such as hepatic lipase, and apolipoprotein E (Klaver et al., 1998; Souied et al., 1998; Baird et al., 2004; Zareparsi et al., 2004; Neale et al., 2010), in immune response, including CD36 and Toll-like receptor-3 and -4 (Zareparsi et al., 2005; Yang et al., 2008), and in retinal function adenosine 5′-triphosphate-binding cassette transporter 4 (ABCA4) (Fritsche et al., 2012; Wu et al., 2015), were also associated with AMD risk.

Despite the strong genetic link, these identified AMD-associated genes only demonstrate susceptibility without proven causality, in contrast to monogenic diseases where a single mutation is sufficient. Given the multi-factorial and complex nature of AMD and its late-in-life onset, one may speculate that genetic risk factors may interact with environmental/lifestyle factors that may precipitate the cellular damage under oxidative stress which accumulates with age, resulting in the risk of AMD development and progression.

Interaction of genetic-oxidative stress response may suggest different levels of genetic susceptibility to oxidative damage

Even with the tremendous effort in investigating AMD genetics, genetic variations alone do not accurately predict AMD progression, as highlighted by monozygotic twin studies, where smoking and dietary nutrient intake dictates disease progression along aging (Seddon et al., 2011). This suggests a significant contribution of environmental factors-induced oxidative stress which may function as modifiers, and also the possibility of gene-environmental interaction determining in part the genetic susceptibilities. Oxidative stress may invoke innate immunity in the eye, through the binding of CFH with malondialdehyde, a common lipid peroxidation product in drusen (Weismann et al., 2011; Alic et al., 2020). The AMD-associated CFH variant H402 strongly reduces its binding affinity to malondialdehyde, suggesting a mechanistic link (Weismann et al., 2011). Similarly, the strong interaction of genetics, including CFH and ARMS2, with antioxidant nutritional supplementation (Vavvas et al., 2018), further suggests a genetics-oxidative stress link that may modify AMD risk.

Additional AMD-associated genes encode proteins comprising the redox scavenging machinery in the eye, such as mitochondria, the main source of energy and ROS generation in RPE. Polymorphism in mitochondrial DNA of MTND2*LHON4917G (4917G) and complex-1 subunit is remarkably associated with AMD development (Canter et al., 2008; SanGiovanni et al., 2009), as well as mitochondrial antioxidant enzymes (Mrowicka et al., 2017), suggesting that compromised antioxidant system prevails the path for RPE degeneration. Additional studies uncovering the link between genetic and environmental factors pertaining to oxidative stress response will provide further insights on AMD pathogenesis.

Enhancing Antioxidant Capacity in Retinal Pigment Epithelium Is a Promising Strategy to Protect against Age-Related Macular Degeneration Progression

Dietary intake of antioxidant vitamins for AMD is endorsed by AREDS & AREDS2 studies

In 1988, a small randomized clinical trial reported for the first time that oral supplementation with zinc, a trace element and essential micro-nutrient, can reduce AMD progression (Newsome et al., 1988). Given the role of zinc in antioxidant response, that trial motivated the use of antioxidant supplementation to protect against AMD development or progression. Later, the beneficial role of antioxidant vitamins in AMD was supported by two large clinical trials: Age-Related Eye Disease Study (AREDS) & Age-Related Eye Disease Study 2 (AREDS2).

Given the chronic nature of AMD, to evaluate whether long-term administration of antioxidants and other dietary supplements may exert greater visual protection, National Eye Institute sponsored AREDS with 3640 individuals for an average of 6.3 years between 1992 and 2001 in the USA, with two primary objectives: first, to investigate the natural course of AMD and age-related cataracts; second, to assess the effects of antioxidants i.e., vitamins C, E, beta-carotene, and zinc on AMD. Original AREDS formula consists of vitamin C (500 mg), vitamin E (400 IU), beta-carotene (15 mg), zinc as zinc oxide (80 mg), and copper as cupric oxide (2 mg) (AREDS, 2001). AREDS showed that nutritional supplementation with antioxidants (vitamins C, E, and beta-carotene) and zinc are protective and reduce the risk of progression to late AMD by 25% in 5 years, by scavenging ROS to ameliorate oxidative stress (AREDS, 2001). In a follow-up study, the beneficial effects of a long-term (10 years) AREDS formulation diet (5 years after the trial ended) persisted in successfully delaying the progression of advanced neovascular AMD in patients with AMD in one eye, but not for the central GA (Chew et al., 2013). AREDS2 study further improved the formula with lutein (10 mg) and zeaxanthin (2 mg), both carotenoids, replacing beta-carotene, to eliminate the concern of beta-carotene in increased lung cancer risk (Chew et al., 2012; AREDS2, 2013). The AREDS2 formula showed incremental protection in intermediate-stage AMD patients compared with the original formula, further confirming the effects of antioxidant vitamins (AREDS2, 2013). Currently, the AREDS2 formula is the only scientifically proven intervention that can delay the progression of intermediate- or late-stage AMD, despite it being non-effective for people with no AMD or early AMD. However, another study has reported varying patient responses (SanGiovanni et al., 2007), potentially reflecting intricate interaction between patient genetics and nutrient intake that warrants further investigation.

Consideration of genetics-nutrient interaction is needed

Since AREDS, the possible influence of the genotype over the AREDS formula treatment response was explored by numerous studies, with findings both supporting and rejecting the idea. Early studies by Awh et al. (2013, 2015) have supported the pharmacogenomics-based AREDS supplement selection in AMD patients. They suggested that the use of zinc and antioxidant should be separated based on specific risk or non-risk allele (CFH/ARMS-2) in patients, with zinc only for those with non-CFH risk, 1–2 risk allele for ARMS-2, and in contrast antioxidant only for those with no-risk ARMS-2 and 1–2 risk allele for CFH (Awh et al., 2013; Awh et al., 2015). These claims were refuted by a separate analysis of AREDS participants by Chew et al., suggesting that AREDS supplements exert similar levels of protection against AMD risk among all genotype groups without any specific influence by CFH risk allele (Chew et al., 2014b), rejecting the need of genetic testing-based ARDES formulation assigning altogether (Chew et al., 2015).

Other later studies found a positive association between AREDS supplement-derived AMD protection and allele variations. For instance, in neovascular AMD patients, AREDS supplements resulted in a lower risk of AMD progression to an advanced stage in high-risk ARMS2 (TT) and non-risk CFH allele (TT) group, but not in CFH high-risk allele (CC) group, yet similar results were not seen in GA (Seddon et al., 2016; Vavvas et al., 2018). On the contrary, AREDS supplement accelerated the rate of AMD progression in high-risk CFH genotype (Vavvas et al., 2018); however, these results were subjected to debate as many other studies, both from AREDS reports and independent analysis of AREDS cohorts, have rejected the genotype-dependent association between AMD progression and AREDS formulation (Assel et al., 2018; van Asten et al., 2019).

Given the debate, more studies will be needed to be able to draw definitive conclusions. Yet consideration of the genetics-nutrient interaction is still warranted, particularly considering the use of supplements in different races and ethnic groups with diverse genetic backgrounds. The occurrence of late AMD is significantly higher in the white population compared to other races in North America (Schachat et al., 1995; Leske et al., 2004; Varma et al., 2004; Restrepo et al., 2014), and they were the primary participants in both US-based AREDS trials. In view of the different dietary intake and genetic makeup in other countries, data related to the nutritional effects on other ethnic populations is still much lacking. This outcome, while await future studies, would have significance on the impact of genetic-nutrient interaction. Nevertheless, current insights obtained have greatly promoted interest in experimental and translational investigations on targeting oxidative stress as a potential therapeutic strategy to protect against AMD in animal models.

Experimental Studies on Oxidative Stress and Cellular Defense: Nuclear Factor Erythroid-2-Related Factor 2 as a Key Antioxidant Enzyme Regulator

NRF2 is a central transcriptional regulator of antioxidant enzymes

Transcriptional regulation of many redox pathway proteins is moderated by NRF2 (encoded by the nfe2l2 gene), a master transcriptional regulator of key antioxidant enzymes. In physiological conditions, NRF2 is sequestered by Kelch-like ECH-associated protein 1 (KEAP1) and negatively regulated through ubiquitination and proteasomal degradation (McMahon et al., 2003). Excess ROS triggers a conformational modification in KEAP1, resulting in the release and subsequent nuclear translocation of NRF2, which then binds to MAF protein and an antioxidant response element on the promoter site of target genes to induce their transcription. These include direct antioxidant enzymes, such as SOD1&2 and catalase, which directly neutralize superoxide and hydrogen peroxide (H2O2), as well as phase 2 enzymes regulating glutathione and thioredoxin levels, and xenobiotic enzymes, e.g. glutathione peroxidase, heme oxygenase 1&2, and NADPH quinine oxidoreductase 1 (Figure 3). Through regulating these antioxidant defense enzymes, NRF2 mediates the redox balance in many cell types including RPE, which has been extensively studied in preclinical animal models of AMD.

Figure 3:
A cartoon illustration of NRF2 signaling.Under physiological conditions, NRF2 is sequestered and negatively regulated by KEAP1 through ubiquitination and proteasomal degradation (left panel). The presence of ROS and oxidative stress triggers conformational modifications in KEAP1, thereby inhibiting its binding to NRF2, inducing its release and nuclear translocation that enables its binding to MAF protein and an ARE on the promoter of its target genes encoding antioxidant enzymes, such as SOD1&2 and catalase, in addition to other phase-2 detoxifying enzymes (right panel). ARE: Antioxidant response element; CUL3: adaptor component Cullin 3; KEAP1: Kelch-like ECH-associated protein 1; MAF: musculoaponeurotic fibrosarcoma oncogene homolog; NRF2: nuclear factor erythroid-2-related factor 2; RBX1: RING box protein 1; RPE: retinal pigment epithelium; SOD: superoxide dismutase. Created with BioRender.com.

Experimental studies on AMD highlight the role of NRF2 in RPE oxidative defense

While there are no perfect animal models of AMD that reproduce all pathological features in humans, experimental studies have benefited much from animal models showing key features of AMD-like pathologies. These include inflammation (Cfh–/– and/or Cx3cr1–/– mice) (Coffey et al., 2007; Combadiere et al., 2007), impaired lipid function (transgenic ApoE4 with high-fat diet, and macrophage-specific ABCA1 and ABCG1 deficiency) (Ramkumar et al., 2010; Ban et al., 2018), and oxidative stress and defense (Sod1–/–, Nrf2–/–, AhR–/–, or carboxyethylpyrrole immunization) (Imamura et al., 2006; Hollyfield et al., 2008; Ramkumar et al., 2010; Zhao et al., 2011; Hu et al., 2013), each displaying some features of AMD (Ramkumar et al., 2010; Pennesi et al., 2012). For example, Sod1–/– mice (Imamura et al., 2006), lacking Cu, Zn-superoxide dismutase-1 (Sod1), a critical enzyme involved in removing superoxide radicals, and hence important for oxidative stress response, exhibited increased drusen with age (from 6–18 months), thickened Bruch’s membrane, disrupted RPE integrity and choroidal neovascularization at 10–12 months of age, with reduced ERG waves, and progressive retinal cell death (Hashizume et al., 2008). In addition, systemic SOD2 knockdown mice also exhibited RPE degeneration and photoreceptor disorganization (Kasahara et al., 2005; Justilien et al., 2007), and RPE-specific deletion of SOD2 led to RPE and photoreceptor metabolic alteration, dysregulation, and dysfunction (Brown et al., 2019). Moreover, sodium-iodate-induced RPE oxidative damage is commonly used as an acute animal model to investigate mechanisms of oxidative damage in RPE (Franco et al., 2009). Together these animal models highlight the role of antioxidant enzymes and oxidative damage in the development of RPE degeneration and AMD-like features.

Given the central role of NRF2 in antioxidant enzyme regulation, a potential role of NRF2 in RPE degeneration is widely investigated. In aging RPE, induction of NRF2 signaling is compromised with oxidative stress (Sachdeva et al., 2014). Nrf2–/– mice developed AMD-like ocular pathologies including sub-RPE deposits with RPE degeneration (Zhao et al., 2011) and these degenerative events accelerated after mild white light exposure (Wang et al., 2019) and increased cigarette smoke-induced RPE damage (Cano et al., 2010), suggesting a key role of NRF2 in RPE oxidative damage and potentially AMD development. In addition, Nrf2–/– mice also developed age-related cataracts (Rowan et al., 2021), highlighting the importance of NRF2-dependent antioxidant signaling in aging eyes. Recently, NRF2 deficiency was also found to impact the mitochondrial antioxidant response by influencing the pentose phosphate pathway in RPE (Cano et al., 2021). Global double knockout of NRF-2 and PGC-1α (peroxisome proliferator-activated receptor gamma coactivator-1 alpha) in mice resulted in significant age-dependent RPE degeneration with mitochondrial damage as well as increased expression of inflammatory and autophagy markers mimicking dry-AMD pathologies (Felszeghy et al., 2019). In addition, a low glycemic diet protected Nrf2–/– mice against AMD-related pathologies, via enhanced antiglycation and anti-oxidative machinery (Rowan et al., 2020). These studies established a pivotal role of NRF2 in RPE and AMD pathophysiology (Cai et al., 2021), yet transcription of NRF2 itself can be subjected to regulation by other factors involved in redox sensing and protection.

REV-ERBα: a newly discovered transcriptional regulator of NRF2 and antioxidant defense in RPE

One such example of NRF2 regulators is REV-ERBα, a redox-sensitive nuclear transcription factor primarily involved in circadian rhythm regulation and metabolism. REV-ERB is a family of redox-sensing nuclear receptors functioning as ligand (heme)-sensitive transcriptional repressors (Raghuram et al., 2007; Kojetin and Burris, 2014; Brown et al., 2018) and a critical sensor of the tissue redox environment in modulating cellular oxidative capacity (Pardee et al., 2009; Gupta and Ragsdale, 2011; Ragsdale et al., 2012; Woldt et al., 2013; Carter and Ragsdale, 2014). A dynamic thiol-disulfide mechanism regulates their ligand binding affinity in a redox-sensitive (thiol-disulfide exchange) manner (Pardee et al., 2009; Ragsdale et al., 2012; Woldt et al., 2013; Carter and Ragsdale, 2014). REV-ERB at a reduced (dithiol) state binds ligand with much higher affinity (5-fold) than the oxidized disulfide state (Gupta and Ragsdale, 2011); therefore, oxidative stress acts through this thiol-disulfide redox switch to lead to oxidation of cysteine(s) residues in REV-ERB, thus causing ligand dissociation to dampen the receptor’s transcriptional activity.

REV-ERBα (also known as Nr1d1) is an essential regulator of many biological processes including circadian rhythm, lipid and glucose metabolism, macrophage inflammatory response, and mitochondrial biosynthesis (Cho et al., 2012; Solt et al., 2012; Lam et al., 2013; Woldt et al., 2013; Zhang et al., 2015b), some of which are highly relevant to RPE health and AMD. Ligand binding to REV-ERBα determines its interaction with other transcriptional co-factors (NCoR and HDAC3) (Zhang et al., 2015b). Upon binding to the specific DNA response element (RORE) in the regulatory region of the target genes, these co-factors together mediate the expression of target genes, particularly circadian clock genes (Woldt et al., 2013; Zhang et al., 2015b). However, for metabolic target gene expression, it was suggested that REV-ERBα recruits co-repressors, such as HDAC3, which tethers REV-ERBα with other tissue lineage-specific transcription factors to mediate the expression of specific metabolic genes tailoring to the need of specific tissues (Woldt et al., 2013; Butler and Burris, 2015; Zhang et al., 2015b).

In the eye, shRNA-targeted knockdown of Rev-erbα in developing mouse eye with sub-retinal shNR1D1 injection led to pan-retinal spotting with decreased visual function (Mollema et al., 2011). REV-ERBα regulates retinal gene transcription in conjunction with NR2E3 (Mollema et al., 2011), and also retinal visual processing and light sensitivity (Ait-Hmyed Hakkari et al., 2016), as well as RPE circadian genes (Milicevic et al., 2019). Genetic overexpression of REV-ERBα as a modifier gene rescues Nr2e3-associated retinal degeneration in rd7 mice (Cruz et al., 2014). Rev-erbα–/– mice have increased numbers of intrinsically photosensitive retinal ganglion cells and demonstrate increased light sensitivity, suggesting a role of REV-ERBα in retinal visual processing (Ait-Hmyed Hakkari et al., 2016).

Recently, we found that REV-ERBα is a direct transcriptional regulator of NRF2 (Huang et al., 2022). Both systemic and RPE cell-specific deficiencies of REV-ERBα result in the downregulation of NRF2 and its target antioxidant enzymes, SOD1 and catalase. More importantly, REV-ERBα-deficient mice exhibit age-dependent ocular pathologies including fundus lesions, thickened Bruch’s membrane, sub-retinal drusenoid deposits, RPE degeneration, and photoreceptor disorganization, suggesting that Rev-erbα–/– mice represent a new animal model of dry AMD. In an oxidative stress-exposed (NaIO3) model of RPE damage, REV-ERBα-deficient mice are more susceptible to RPE oxidative damage, whereas treatment with REV-ERBα agonist, SR0009 protects against RPE damage with upregulation of NRF2, SOD1, and catalase (Huang et al., 2022). These findings together suggest a protective role of REV-ERBα against RPE degeneration in response to oxidative stress and that targeting REV-ERBα may help protect RPE in dry AMD (Figure 4). In other studies, REV-ERBα agonists were found to be cytoprotective in general, and effective in promoting skeleton muscle cell survival, suppressing diabetes-induced metabolic changes, and atherosclerosis (Solt et al., 2012; Woldt et al., 2013; Sitaula et al., 2015; Ait-Hmyed Hakkari et al., 2016) and promoting the targeted killing of cancer cells (Sulli et al., 2018). REV-ERBα agonists may thus serve as attractive drug target candidates for developing therapeutics for not only AMD but also other diseases.

Figure 4:
A graphical illustration of REV-ERBα-dependent transcriptional regulation of antioxidant response through NRF2 in RPE relevant to AMD pathophysiology.REV-ERBα is a redox-sensitive ligand-dependent receptor and transcriptional factor. Aging (and resultant oxidative stress) leads to the decline of REV-ERBα in RPE. REV-ERBα binds directly to the RORE element in the promoter region of the NRF2 gene (Nfe2l2) to promote its transcription and further increases the levels of NRF2-mediated antioxidant enzymes such as SOD1 and catalase. Genetic deletion of REV-ERBα in mice decreases transcription of NRF2 and its downstream target genes in RPE, thereby leading to oxidative damage, RPE dysfunction and degeneration, with subretinal drusen-like lipid deposit. Activation of REV-ERBα represents a novel therapeutic approach to enhance the NRF2-mediated antioxidant defense system and protect against RPE damage associated with dry AMD. AMD: Age-related macular degeneration; NRF2: nuclear factor erythroid-2-related factor 2; RORE: ROR response element; RPE: retinal pigment epithelium; SOD1: superoxide dismutase-1. Cartoon in the right panel box was created with BioRender.com.

Promoting NRF2 to counter oxidative damage in RPE

Because of its essential role in sensing redox stress, NRF2 has become a molecular target for eye disease protection and particularly RPE protection against oxidative stress. Indeed, RPE-specific NRF-2 overexpression via adeno-associated virus vector delivery preserves RPE morphology and cone photoreceptors in mouse models of retinitis pigmentosa (Wu et al., 2021). In addition, intravitreal delivery of adeno-associated virus containing an NRF2-derived peptide protected RPE and partially restored visual function in the NaIO3-induced RPE oxidative injury model by inducing antioxidant pathway and dampening inflammation (Ildefonso et al., 2016). Moreover, overexpression of NRF2 also protects retinal neurons including photoreceptors and retinal ganglion cells in rodent models of photoreceptor degeneration and nerve crush, more than viral delivery of SOD2 and catalase (Xiong et al., 2015), suggesting the promise of targeting NRF2 in protecting retinal neurons and RPE against oxidative damage.

Many studies explored both upstream and downstream redox regulatory mechanisms of NRF2 in RPE using various in vitro and in vivo model systems. These models rely on the induction of oxidative stress by free radical generating substances, such as sodium iodate-induced oxidative stress model (Han et al., 2020; Tang et al., 2021), white/blue light exposure (Liu et al., 2021; Xie et al., 2021), cigarette smoke extract exposure (Huang et al., 2015) and H2O2-induced oxidative damage model (Du et al., 2020; Ma et al., 2021; You et al., 2021). Together these work in cell culture (Huang et al., 2015; Du et al., 2020; Liu et al., 2021; Ma et al., 2021; Tang et al., 2021; Xie et al., 2021; You et al., 2021) and animal-based models (Han et al., 2020; Xie et al., 2021) discover additional regulators of NRF2 signaling, just like REV-ERBα. For instance, X box-binding protein 1, an endoplasmic reticulum stress-inducible transcription factor (Huang et al., 2015; Chen et al., 2018) and microphthalmia-associated transcription factor (Han et al., 2020) also act as a novel upstream modulator of NRF2 against oxidative stress damage in RPE. Targeting microRNA is also emerging as an effective strategy to modulate upstream NRF2 signaling cascade to counter oxidative stress in RPE, for instance, targeting cullin 3 by miR-601 against H2O2-induced stress (Chen et al., 2019), or Keap1 by microRNA-141 (Cheng et al., 2017) and miR-125b (Liu et al., 2022).

Other additional studies focused on the effects of pharmacological modulators with chemical compounds to influence NRF2 signaling in RPE preservation. Smoking, in particular, impairs NRF2 signaling in aging RPE cells (Suzuki et al., 2008; Wang et al., 2014). Exposure of cigarette smoke extract to NRF2-deficient RPE cells exhibited elevated oxidative stress due to decreased expression of IDH2 and pentose phosphate pathway genes (Cano et al., 2021). In the blue light-driven oxidative stress model of RPE cell culture and BALB/c mice, treatment with lipoxin A4, an antioxidant lipid, reduces the blue light-driven oxidative stress by upregulating heme oxygenase-1 (HO-1) and nuclear translocation of NRF-2 (Xie et al., 2021). In the H2O2-induced oxidative damage model, RPE cells were pre-treated with different chemical compounds: including ferrerol, a rhododendron-derived NRF2 activator (Ma et al., 2021), catalpol, an iridoid glucoside from Rehmannia glutinosa Libosch plant root dry extract (You et al., 2021), and phillyrin, extracted from Chinese herbaceous plant Forsythia suspense (Thunb.) Vah fruits with anti-inflammatory and antioxidant properties (Du et al., 2020). Together these compounds revealed NRF2 activating potential and subsequent elevation of its downstream antioxidant enzymes: catalase, HO-1, NADPH dehydrogenase, SOD and increased glutathione, and hence inhibition of endogenous and exogenous apoptotic pathways. In these studies, inhibition of Keap1/NRF2 protein complex formation, nuclear translocation of NRF2, and elevation in HO-1 expression levels were consistently shown among many oxidative stress models, suggesting that HO-1 upregulation by NRF2 is one of the key hierarchical axis against the oxidative stress in RPE (Shivarudrappa and Ponesakki, 2020; Chen et al., 2021; Ma et al., 2021; Tang et al., 2021; You et al., 2021; Zhao et al., 2021), in addition to other NRF2-mediated antioxidant enzymes. Together these studies demonstrated the translational potential of promoting activation and expression of NRF2, its upstream regulators, and downstream target enzymes, to counter oxidative stress-induced RPE damage.


As dry AMD can progress to neovascular AMD, treating patients at the early stage of dry AMD could potentially prevent or slow down the disease progression to the late devastating stages of GA and CNV. Protection of RPE against oxidative damage is vital for slowing AMD progression and preservation of vision. In recent years, stem cell transplantation and RPE patch transplantation have shown great promise in patients with AMD (Mandai et al., 2017; Yang et al., 2022), yet safety and efficacy will await more studies. Preventive measures to halt or slow down AMD progression, however, are still very negligible. On that note, long-term dietary intake of antioxidant supplements suggested by AREDS & AREDS2 studies significantly reduces the progression to late-stage AMD, highlighting the importance of oxidative damage in AMD pathogenesis. However, there are no current pharmacological approaches to slow or stop dry AMD progression or curb oxidative injury in RPE.

With both human genetics and experimental studies strongly supporting the role of redox imbalance in RPE oxidative injury during aging, limiting oxidative stress may thus act as a preventive and therapeutic measure against RPE oxidative damage and degeneration. Key transcriptional regulatory factors of antioxidant pathways such as NRF2 can be targeted to drive the cascade of synchronous expression of various antioxidant enzymes and pro-survival proteins to protect RPE in AMD. Identification of novel upstream regulators of NRF2, such as REV-ERBα, adds new ways to modulate antioxidant pathways and enhance RPE antioxidant self-defense system. Future work can be directed towards the revelation of the intertwined interaction of immune and inflammatory, metabolic, and antioxidant pathways in AMD. That will further improve our understanding of AMD pathogenesis and allow the development of new effective therapeutic strategies to counter RPE oxidative damage for disease management.

Author contributions:MM and JC conceived and wrote the review; MM and CHL prepared the table and figures; all authors critically reviewed, edited and approved the manuscript.

Conflicts of interest:The authors declare no conflicts of interest. No conflicts of interest exist between Mass Lions Eye Research Fund Inc. and publication of the manuscript.

Data availability statement:The data are available from the corresponding author on reasonable request.

Open peer reviewer:Lawrence J. Rizzolo, Yale University School of Medicine, USA.

Additional file:Open peer review report 1.


P-Reviewers: Rizzolo LJ, Zhang H; C-Editors: Zhao M, Liu WJ, Qiu Y; T-Editor: Jia Y


1. Age-Related Eye Disease Study Research Group [AREDS] 2001 A randomized, placebo-controlled, clinical trial of high-dose supplementation with vitamins C and E, beta carotene, and zinc for age-related macular degeneration and vision loss:AREDS report No. 8. Arch Ophthalmol 119:1417–1436.
2. Age-Related Eye Disease Study 2 Research Group [AREDS2] 2013 Lutein +zeaxanthin and omega-3 fatty acids for age-related macular degeneration:the Age-Related Eye Disease Study 2 (AREDS2) randomized clinical trial. JAMA 309:2005–2015.
3. Ait-Hmyed Hakkari O, Acar N, Savier E, Spinnhirny P, Bennis M, Felder-Schmittbuhl MP, Mendoza J, Hicks D 2016 Rev-Erbαmodulates retinal visual processing and behavioral responses to light. FASEB J 30:3690–3701.
4. Alic L, Papac-Milicevic N, Czamara D, Rudnick RB, Ozsvar-Kozma M, Hartmann A, Gurbisz M, Hoermann G, Haslinger-Hutter S, Zipfel PF, Skerka C, Binder EB, Binder CJ 2020 A genome-wide association study identifies key modulators of complement factor H binding to malondialdehyde-epitopes. Proc Natl Acad Sci U S A 117:9942–9951.
5. Assel MJ, Li F, Wang Y, Allen AS, Baggerly KA, Vickers AJ 2018 Genetic polymorphisms of CFH and ARMS2 do not predict response to antioxidants and zinc in patients with age-related macular degeneration:independent statistical evaluations of data from the age-related eye disease study. Ophthalmology 125:391–397.
6. Awh CC, Lane AM, Hawken S, Zanke B, Kim IK 2013 CFH and ARMS2 genetic polymorphisms predict response to antioxidants and zinc in patients with age-related macular degeneration. Ophthalmology 120:2317–2323.
7. Awh CC, Hawken S, Zanke BW 2015 Treatment response to antioxidants and zinc based on CFH and ARMS2 genetic risk allele number in the age-related eye disease study. Ophthalmology 122:162–169.
8. Baird PN, Guida E, Chu DT, Vu HT, Guymer RH 2004 The epsilon2 and epsilon4 alleles of the apolipoprotein gene are associated with age-related macular degeneration. Invest Ophthalmol Vis Sci 45:1311–1315.
9. Ban N, Lee TJ, Sene A, Choudhary M, Lekwuwa M, Dong Z, Santeford A, Lin JB, Malek G, Ory DS, Apte RS 2018 Impaired monocyte cholesterol clearance initiates age-related retinal degeneration and vision loss. JCI Insight 3:e120824.
10. Beatty S, Koh H, Phil M, Henson D, Boulton M 2000 The role of oxidative stress in the pathogenesis of age-related macular degeneration. Surv Ophthalmol 45:115–134.
11. Bertram KM, Baglole CJ, Phipps RP, Libby RT 2009 Molecular regulation of cigarette smoke induced-oxidative stress in human retinal pigment epithelial cells:implications for age-related macular degeneration. Am J Physiol Cell Physiol 297:C1200–1210.
12. Bird A 2021 Role of retinal pigment epithelium in age-related macular disease:a systematic review. Br J Ophthalmol 105:1469–1474.
13. Bressler NM 2002 Early detection and treatment of neovascular age-related macular degeneration. J Am Board Fam Pract 15:142–152.
14. Bressler SB, Muñoz B, Solomon SD, West SK 2008 Racial differences in the prevalence of age-related macular degeneration:the Salisbury Eye Evaluation (SEE) Project. Arch Ophthalmol 126:241–245.
15. Brown EE, Lewin AS, Ash JD 2018 Mitochondria:potential targets for protection in age-related macular degeneration. Adv Exp Med Biol 1074:11–17.
16. Brown EE, DeWeerd AJ, Ildefonso CJ, Lewin AS, Ash JD 2019 Mitochondrial oxidative stress in the retinal pigment epithelium (RPE) led to metabolic dysfunction in both the RPE and retinal photoreceptors. Redox Biol 24:101201.
17. Butler AA, Burris TP 2015 Segregation of clock and non-clock regulatory functions of REV-ERB. Cell Metab 22:197–198.
18. Cadenas E, Davies KJA 2000 Mitochondrial free radical generation, oxidative stress, and aging. Free Radic Biol Med 29:222–230.
19. Cai J, Nelson KC, Wu M, Sternberg P Jr, Jones DP 2000 Oxidative damage and protection of the RPE. Prog Retin Eye Res 19:205–221.
20. Cai J, Zhang H, Zhang YF, Zhou Z, Wu S 2019 MicroRNA-29 enhances autophagy and cleanses exogenous mutant αB-crystallin in retinal pigment epithelial cells. Exp Cell Res 374:231–248.
21. Cai ZY, Fu MD, Liu K, Duan XC 2021 Therapeutic effect of Keap1-Nrf2-ARE pathway-related drugs on age-related eye diseases through anti-oxidative stress. Int J Ophthalmol 14:1260–1273.
22. Cano M, Datta S, Wang L, Liu T, Flores-Bellver M, Sachdeva M, Sinha D, Handa JT 2021 Nrf2 deficiency decreases NADPH from impaired IDH shuttle and pentose phosphate pathway in retinal pigmented epithelial cells to magnify oxidative stress-induced mitochondrial dysfunction. Aging Cell 20:e13444.
23. Cano M, Thimmalappula R, Fujihara M, Nagai N, Sporn M, Wang AL, Neufeld AH, Biswal S, Handa JT 2010 Cigarette smoking, oxidative stress, the anti-oxidant response through Nrf2 signaling, and age-related macular degeneration. Vision Res 50:652–664.
24. Canter JA, Olson LM, Spencer K, Schnetz-Boutaud N, Anderson B, Hauser MA, Schmidt S, Postel EA, Agarwal A, Pericak-Vance MA, Sternberg P Jr, Haines JL 2008 Mitochondrial DNA polymorphism A4917G is independently associated with age-related macular degeneration. PLoS One 3:e2091.
25. Carter EL, Ragsdale SW 2014 Modulation of nuclear receptor function by cellular redox poise. J Inorg Biochem 133:92–103.
26. Chen C, Zhong Y, Wang JJ, Yu Q, Plafker K, Plafker S, Zhang SX 2018 Regulation of Nrf2 by X box-binding protein 1 in retinal pigment epithelium. Front Genet 9:658.
27. Chen W, Ye Y, Wu Z, Lin J, Wang Y, Ding Q, Yang X, Yang W, Lin B, Lin B 2021 Temporary upregulation of Nrf2 by naringenin alleviates oxidative damage in the retina and ARPE-19 cells. Oxid Med Cell Longev 2021:4053276.
28. Chen ZJ, Rong L, Huang D, Jiang Q 2019 Targeting cullin 3 by miR-601 activates Nrf2 signaling to protect retinal pigment epithelium cells from hydrogen peroxide. Biochem Biophys Res Commun 515:679–687.
29. Cheng LB, Li KR, Yi N, Li XM, Wang F, Xue B, Pan YS, Yao J, Jiang Q, Wu ZF 2017 miRNA-141 attenuates UV-induced oxidative stress via activating Keap1-Nrf2 signaling in human retinal pigment epithelium cells and retinal ganglion cells. Oncotarget 8:13186–13194.
30. Chew EY, Clemons T, SanGiovanni JP, Danis R, Domalpally A, McBee W, Sperduto R, Ferris FL 2012 The Age-Related Eye Disease Study 2 (AREDS2):study design and baseline characteristics (AREDS2 report number 1). Ophthalmology 119:2282–2289.
31. Chew EY, Clemons TE, Agrón E, Sperduto RD, Sangiovanni JP, Kurinij N, Davis MD 2013 Long-term effects of vitamins C and E, β-carotene, and zinc on age-related macular degeneration:AREDS report no. 35. Ophthalmology 120:1604–1611.
32. Chew EY, Clemons TE, Agrón E, Sperduto RD, Sangiovanni JP, Davis MD, Ferris FL 3rd 2014a Ten-year follow-up of age-related macular degeneration in the age-related eye disease study:AREDS report no. 36. JAMA Ophthalmol 132:272–277.
33. Chew EY, Klein ML, Clemons TE, Agrón E, Ratnapriya R, Edwards AO, Fritsche LG, Swaroop A, Abecasis GR 2014b No clinically significant association between CFH and ARMS2 genotypes and response to nutritional supplements:AREDS Report Number 38. Ophthalmology 121:2173–2180.
34. Chew EY, Klein ML, Clemons TE, Agrón E, Abecasis GR 2015 Genetic testing in persons with age-related macular degeneration and the use of the AREDS supplements:to test or not to test?. Ophthalmology 122:212–215.
35. Chiu CJ, Chang ML, Zhang FF, Li T, Gensler G, Schleicher M, Taylor A 2014 The relationship of major American dietary patterns to age-related macular degeneration. Am J Ophthalmol 158:118–127.
36. Cho H, Zhao X, Hatori M, Yu RT, Barish GD, Lam MT, Chong LW, DiTacchio L, Atkins AR, Glass CK, Liddle C, Auwerx J, Downes M, Panda S, Evans RM 2012 Regulation of circadian behaviour and metabolism by REV-ERB-alpha and REV-ERB-beta. Nature 485:123–127.
37. Coffey PJ, Gias C, McDermott CJ, Lundh P, Pickering MC, Sethi C, Bird A, Fitzke FW, Maass A, Chen LL, Holder GE, Luthert PJ, Salt TE, Moss SE, Greenwood J 2007 Complement factor H deficiency in aged mice causes retinal abnormalities and visual dysfunction. Proc Natl Acad Sci U S A 104:16651–16656.
38. Combadiere C, Feumi C, Raoul W, Keller N, Rodero M, Pezard A, Lavalette S, Houssier M, Jonet L, Picard E, Debre P, Sirinyan M, Deterre P, Ferroukhi T, Cohen SY, Chauvaud D, Jeanny JC, Chemtob S, Behar-Cohen F, Sennlaub F 2007 CX3CR1-dependent subretinal microglia cell accumulation is associated with cardinal features of age-related macular degeneration. J Clin Invest 117:2920–2928.
39. Cruickshanks KJ, Klein R, Klein BEK 1993 Sunlight and age-related macular degeneration:the beaver dam eye study. Arch Ophthalmol 111:514–518.
40. Cruz NM, Yuan Y, Leehy BD, Baid R, Kompella U, DeAngelis MM, Escher P, Haider NB 2014 Modifier genes as therapeutics:the nuclear hormone receptor Rev Erb alpha (Nr1d1) rescues Nr2e3 associated retinal disease. PLoS One 9:e87942.
41. Curcio CA 2018 Soft drusen in age-related macular degeneration:biology and targeting via the oil spill strategies. Invest Ophthalmol Vis Sci 59:Amd160–amd181.
42. Datta S, Cano M, Ebrahimi K, Wang L, Handa JT 2017 The impact of oxidative stress and inflammation on RPE degeneration in non-neovascular AMD. Prog Retin Eye Res 60:201–218.
43. De Jong PT, Klaver CC, Wolfs RC, Assink JJ, Hofman A 1997 Familial aggregation of age-related maculopathy. Am J Ophthalmol 124:862–863.
44. de Jong PT 2016 A historical analysis of the quest for the origins of aging macula disorder, the tissues involved, and its terminology. Ophthalmol Eye Dis 8:5–14.
45. Dithmar S, Sharara NA, Curcio CA, Le NA, Zhang Y, Brown S, Grossniklaus HE 2001 Murine high-fat diet and laser photochemical model of basal deposits in Bruch membrane. Arch Ophthalmol 119:1643–1649.
46. Dosso AA, Bovet J 1992 Monozygotic twin brothers with age-related macular degeneration. Ophthalmologica 205:24–28.
47. Du SW, Palczewski K 2022 MicroRNA regulation of critical retinal pigment epithelial functions. Trends Neurosci 45:78–90.
48. Du Y, You L, Ni B, Sai N, Wang W, Sun M, Xu R, Yao Y, Zhang Z, Qu C, Yin X, Ni J 2020 Phillyrin mitigates apoptosis and oxidative stress in hydrogen peroxide-treated RPE cells through activation of the Nrf2 signaling pathway. Oxid Med Cell Longev 2020:2684672.
49. Edwards AO, Ritter R 3rd, Abel KJ, Manning A, Panhuysen C, Farrer LA 2005 Complement factor H polymorphism and age-related macular degeneration. Science 308:421–424.
50. Evans J 2008 Antioxidant supplements to prevent or slow down the progression of AMD:a systematic review and meta-analysis. Eye (Lond) 22:751–760.
51. Felszeghy S, Viiri J, Paterno JJ, Hyttinen JMT, Koskela A, Chen M, Leinonen H, Tanila H, Kivinen N, Koistinen A, Toropainen E, Amadio M, Smedowski A, Reinisalo M, Winiarczyk M, Mackiewicz J, Mutikainen M, Ruotsalainen AK, Kettunen M, Jokivarsi K, et al. 2019 Loss of NRF-2 and PGC-1αgenes leads to retinal pigment epithelium damage resembling dry age-related macular degeneration. Redox Biol 20:1–12.
52. Ferris FL 3rd, Wilkinson CP, Bird A, Chakravarthy U, Chew E, Csaky K, Sadda SR 2013 Clinical classification of age-related macular degeneration. Ophthalmology 120:844–851.
53. Fields MA, Del Priore LV, Adelman RA, Rizzolo LJ 2020 Interactions of the choroid, Bruch's membrane, retinal pigment epithelium, and neurosensory retina collaborate to form the outer blood-retinal-barrier. Prog Retin Eye Res 76:100803.
54. Fisher CR, Ferrington DA 2018 Perspective on AMD pathobiology:a bioenergetic crisis in the RPE. Invest Ophthalmol Vis Sci 59:Amd41–amd47.
55. Fleckenstein M, Keenan TDL, Guymer RH, Chakravarthy U, Schmitz-Valckenberg S, Klaver CC, Wong WT, Chew EY 2021 Age-related macular degeneration. Nat Rev Dis Primers 7:31.
56. Francis PJ, Hamon SC, Ott J, Weleber RG, Klein ML 2009 Polymorphisms in C2, CFB and C3 are associated with progression to advanced age related macular degeneration associated with visual loss. J Med Genet 46:300–307.
57. Franco LM, Zulliger R, Wolf-Schnurrbusch UE, Katagiri Y, Kaplan HJ, Wolf S, Enzmann V 2009 Decreased visual function after patchy loss of retinal pigment epithelium induced by low-dose sodium iodate. Invest Ophthalmol Vis Sci 50:4004–4010.
58. Friedman DS, O'Colmain BJ, Munoz B, Tomany SC, McCarty C, de Jong PT, Nemesure B, Mitchell P, Kempen J 2004 Prevalence of age-related macular degeneration in the United States. Arch Ophthalmol 122:564–572.
59. Fritsche LG, Fleckenstein M, Fiebig BS, Schmitz-Valckenberg S, Bindewald-Wittich A, Keilhauer CN, Renner AB, Mackensen F, Mößner A, Pauleikhoff D, Adrion C, Mansmann U, Scholl HP, Holz FG, Weber BH 2012 A subgroup of age-related macular degeneration is associated with mono-allelic sequence variants in the ABCA4 gene. Invest Ophthalmol Vis Sci 53:2112–2118.
60. Gehrs KM, Anderson DH, Johnson LV, Hageman GS 2006 Age-related macular degeneration--emerging pathogenetic and therapeutic concepts. Ann Med 38:450–471.
61. Gold B, Merriam JE, Zernant J, Hancox LS, Taiber AJ, Gehrs K, Cramer K, Neel J, Bergeron J, Barile GR, Smith RT, Hageman GS, Dean M, Allikmets R 2006 Variation in factor B (BF) and complement component 2 (C2) genes is associated with age-related macular degeneration. Nat Genet 38:458–462.
62. Golestaneh N, Chu Y, Xiao YY, Stoleru GL, Theos AC 2017 Dysfunctional autophagy in RPE, a contributing factor in age-related macular degeneration. Cell Death Dis 8:e2537.
63. Gounder SS, Kannan S, Devadoss D, Miller CJ, Whitehead KJ, Odelberg SJ, Firpo MA, Paine R 3rd, Hoidal JR, Abel ED, Rajasekaran NS 2012 Impaired transcriptional activity of Nrf2 in age-related myocardial oxidative stress is reversible by moderate exercise training. PLoS One 7:e45697.
64. Grizzard SW, Arnett D, Haag SL 2003 Twin study of age-related macular degeneration. Ophthalmic Epidemiol 10:315–322.
65. Gupta N, Ragsdale SW 2011 Thiol-disulfide redox dependence of heme binding and heme ligand switching in nuclear hormone receptor rev-erb{beta}. J Biol Chem 286:4392–4403.
66. Haddad S, Chen CA, Santangelo SL, Seddon JM 2006 The genetics of age-related macular degeneration:a review of progress to date. Surv Ophthalmol 51:316–363.
67. Hadziahmetovic M, Malek G 2020 Age-related macular degeneration revisited:from pathology and cellular stress to potential therapies. Front Cell Dev Biol 8:612812.
68. Hageman GS, Luthert PJ, Victor Chong NH, Johnson LV, Anderson DH, Mullins RF 2001 An integrated hypothesis that considers drusen as biomarkers of immune-mediated processes at the RPE-Bruch's membrane interface in aging and age-related macular degeneration. Prog Retin Eye Res 20:705–732.
69. Haines JL, Hauser MA, Schmidt S, Scott WK, Olson LM, Gallins P, Spencer KL, Kwan SY, Noureddine M, Gilbert JR, Schnetz-Boutaud N, Agarwal A, Postel EA, Pericak-Vance MA 2005 Complement factor H variant increases the risk of age-related macular degeneration. Science 308:419–421.
70. Hammond CJ, Webster AR, Snieder H, Bird AC, Gilbert CE, Spector TD 2002 Genetic influence on early age-related maculopathy:a twin study. Ophthalmology 109:730–736.
71. Han S, Chen J, Hua J, Hu X, Jian S, Zheng G, Wang J, Li H, Yang J, Hejtmancik JF, Qu J, Ma X, Hou L 2020 MITF protects against oxidative damage-induced retinal degeneration by regulating the NRF2 pathway in the retinal pigment epithelium. Redox Biol 34:101537.
72. Hashizume K, Hirasawa M, Imamura Y, Noda S, Shimizu T, Shinoda K, Kurihara T, Noda K, Ozawa Y, Ishida S, Miyake Y, Shirasawa T, Tsubota K 2008 Retinal dysfunction and progressive retinal cell death in SOD1-deficient mice. Am J Pathol 172:1325–1331.
73. Hollyfield JG, Bonilha VL, Rayborn ME, Yang X, Shadrach KG, Lu L, Ufret RL, Salomon RG, Perez VL 2008 Oxidative damage-induced inflammation initiates age-related macular degeneration. Nat Med 14:194–198.
74. Hu P, Herrmann R, Bednar A, Saloupis P, Dwyer MA, Yang P, Qi X, Thomas RS, Jaffe GJ, Boulton ME, McDonnell DP, Malek G 2013 Aryl hydrocarbon receptor deficiency causes dysregulated cellular matrix metabolism and age-related macular degeneration-like pathology. Proc Natl Acad Sci U S A 110:E4069–4078.
75. Huang C, Wang JJ, Ma JH, Jin C, Yu Q, Zhang SX 2015 Activation of the UPR protects against cigarette smoke-induced RPE apoptosis through up-regulation of Nrf2. J Biol Chem 290:5367–5380.
76. Huang S, Liu CH, Wang Z, Fu Z, Britton WR, Blomfield AK, Kamenecka TM, Dunaief JL, Solt LA, Chen J 2022 REV-ERBαregulates age-related and oxidative stress-induced degeneration in retinal pigment epithelium via NRF2. Redox Biology 51:102261.
77. Hyttinen JMT, Blasiak J, Felszeghy S, Kaarniranta K 2021 MicroRNAs in the regulation of autophagy and their possible use in age-related macular degeneration therapy. Ageing Res Rev 67:101260.
78. Ildefonso CJ, Jaime H, Brown EE, Iwata RL, Ahmed CM, Massengill MT, Biswal MR, Boye SE, Hauswirth WW, Ash JD, Li Q, Lewin AS 2016 Targeting the Nrf2 signaling pathway in the retina with a gene-delivered secretable and cell-penetrating peptide. Invest Ophthalmol Vis Sci 57:372–386.
79. Imamura Y, Noda S, Hashizume K, Shinoda K, Yamaguchi M, Uchiyama S, Shimizu T, Mizushima Y, Shirasawa T, Tsubota K 2006 Drusen, choroidal neovascularization, and retinal pigment epithelium dysfunction in SOD1-deficient mice:a model of age-related macular degeneration. Proc Natl Acad Sci U S A 103:11282–11287.
80. Intartaglia D, Giamundo G, Conte I 2020 The impact of miRNAs in health and disease of retinal pigment epithelium. Front Cell Dev Biol 8:589985.
81. Jia L, Liu Z, Sun L, Miller SS, Ames BN, Cotman CW, Liu J 2007 Acrolein, a toxicant in cigarette smoke, causes oxidative damage and mitochondrial dysfunction in RPE cells:protection by (R)-alpha-lipoic acid. Invest Ophthalmol Vis Sci 48:339–348.
82. Jiang C, Xie P, Sun R, Sun X, Liu G, Ding S, Zhu M, Yan B, Liu Q, Chen X, Zhao C 2018 c-Jun-mediated microRNA-302d-3p induces RPE dedifferentiation by targeting p21(Waf1/Cip1). Cell Death Dis 9:451.
83. Jun S, Datta S, Wang L, Pegany R, Cano M, Handa JT 2019 The impact of lipids, lipid oxidation, and inflammation on AMD, and the potential role of miRNAs on lipid metabolism in the RPE. Exp Eye Res 181:346–355.
84. Justilien V, Pang JJ, Renganathan K, Zhan X, Crabb JW, Kim SR, Sparrow JR, Hauswirth WW, Lewin AS 2007 SOD2 knockdown mouse model of early AMD. Invest Ophthalmol Vis Sci 48:4407–4420.
85. Kaarniranta K, Pawlowska E, Szczepanska J, Jablkowska A, Blasiak J 2019 Role of mitochondrial DNA damage in ROS-mediated pathogenesis of age-related macular degeneration (AMD). Int J Mol Sci 20:2374.
86. Kaarniranta K, Pawlowska E, Szczepanska J, Blasiak J 2020 DICER1 in the pathogenesis of age-related macular degeneration (AMD) - Alu RNA Accumulation versus miRNA Dysregulation. Aging Dis 11:851–862.
87. Kaemmerer E, Schutt F, Krohne TU, Holz FG, Kopitz J 2007 Effects of lipid peroxidation-related protein modifications on RPE lysosomal functions and POS phagocytosis. Invest Ophthalmol Vis Sci 48:1342–1347.
88. Kanda A, Stambolian D, Chen W, Curcio CA, Abecasis GR, Swaroop A 2010 Age-related macular degeneration-associated variants at chromosome 10q26 do not significantly alter ARMS2 and HTRA1 transcript levels in the human retina. Mol Vis 16:1317–1323.
89. Kaneko H, Dridi S, Tarallo V, Gelfand BD, Fowler BJ, Cho WG, Kleinman ME, Ponicsan SL, Hauswirth WW, Chiodo VA, Karikó K, Yoo JW, Lee DK, Hadziahmetovic M, Song Y, Misra S, Chaudhuri G, Buaas FW, Braun RE, Hinton DR, et al. 2011 DICER1 deficit induces Alu RNA toxicity in age-related macular degeneration. Nature 471:325–330.
90. Kasahara E, Lin LR, Ho YS, Reddy VN 2005 SOD2 protects against oxidation-induced apoptosis in mouse retinal pigment epithelium:implications for age-related macular degeneration. Invest Ophthalmol Vis Sci 46:3426–3434.
91. Klaver CC, Wolfs RC, Assink JJ, van Duijn CM, Hofman A, de Jong PT 1998 Genetic risk of age-related maculopathy. Population-based familial aggregation study. Arch Ophthalmol 116:1646–1651.
92. Klein ML, Mauldin WM, Stoumbos VD 1994 Heredity and age-related macular degeneration. Observations in monozygotic twins. Arch Ophthalmol 112:932–937.
93. Klein R, Klein BE, Linton KL 1992 Prevalence of age-related maculopathy. The Beaver Dam Eye Study. Ophthalmology 99:933–943.
94. Klein R, Klein BEK, Linton KLP, DeMets DL 1993 The Beaver Dam Eye Study:The Relation of Age-related Maculopathy to Smoking. Am J Epidemiol 137:190–200.
95. Klein R, Klein BE, Moss SE 1998 Relation of smoking to the incidence of age-related maculopathy. The Beaver Dam Eye Study. Am J Epidemiol 147:103–110.
96. Klein R, Klein BE, Knudtson MD, Wong TY, Cotch MF, Liu K, Burke G, Saad MF, Jacobs DR Jr 2006 Prevalence of age-related macular degeneration in 4 racial/ethnic groups in the multi-ethnic study of atherosclerosis. Ophthalmology 113:373–380.
97. Klein RJ, Zeiss C, Chew EY, Tsai JY, Sackler RS, Haynes C, Henning AK, SanGiovanni JP, Mane SM, Mayne ST, Bracken MB, Ferris FL, Ott J, Barnstable C, Hoh J 2005 Complement factor H polymorphism in age-related macular degeneration. Science 308:385–389.
98. Kojetin DJ, Burris TP 2014 REV-ERB and ROR nuclear receptors as drug targets. Nat Rev Drug Discov 13:197–216.
99. Kuan V, Warwick A, Hingorani A, Tufail A, Cipriani V, Burgess S, Sofat R 2021 Association of smoking, alcohol consumption, blood pressure, body mass index, and glycemic risk factors with age-related macular degeneration:a mendelian randomization study. JAMA Ophthalmol 139:1299–1306.
100. Lam MT, Cho H, Lesch HP, Gosselin D, Heinz S, Tanaka-Oishi Y, Benner C, Kaikkonen MU, Kim AS, Kosaka M, Lee CY, Watt A, Grossman TR, Rosenfeld MG, Evans RM, Glass CK 2013 Rev-Erbs repress macrophage gene expression by inhibiting enhancer-directed transcription. Nature 498:511–515.
101. Lambros ML, Plafker SM 2016 Oxidative stress and the Nrf2 anti-oxidant transcription factor in age-related macular degeneration. Adv Exp Med Biol 854:67–72.
102. LaVail MM 1976 Rod outer segment disk shedding in rat retina:relationship to cyclic lighting. Science 194:1071–1074.
103. Leske MC, Wu SY, Hyman L, Hennis A, Nemesure B, Schachat AP 2004 Four-year incidence of macular changes in the Barbados Eye Studies. Ophthalmology 111:706–711.
104. Liang FQ, Godley BF 2003 Oxidative stress-induced mitochondrial DNA damage in human retinal pigment epithelial cells:a possible mechanism for RPE aging and age-related macular degeneration. Exp Eye Res 76:397–403.
105. Liu CH, Huang S, Britton WR, Chen J 2020 MicroRNAs in vascular eye diseases. Int J Mol Sci 21:649.
106. Liu JX, Ma DY, Zhi XY, Wang MW, Zhao JY, Qin Y 2022 MiR-125b attenuates retinal pigment epithelium oxidative damage via targeting Nrf2/HIF-1αsignal pathway. Exp Cell Res 410:112955.
107. Liu Y, Fiskum G, Schubert D 2002 Generation of reactive oxygen species by the mitochondrial electron transport chain. J Neurochem 80:780–787.
108. Liu Y, Guo Z, Wang S, Liu Y, Wei Y 2021 Fucoxanthin pretreatment ameliorates visible light-induced phagocytosis disruption of RPE cells under a lipid-rich environment via the Nrf2 pathway. Mar Drugs 20:15.
109. Ma N, Yang X, Qi C, Yu Q, Zhu C, Ren H 2021 Farrerol enhances Nrf2-mediated defense mechanisms against hydrogen peroxide-induced oxidative damage in human retinal pigment epithelial cells by activating Akt and MAPK. Oxid Med Cell Longev 2021:8847844.
110. Malek G, Dwyer M, McDonnell D 2012 Exploring the potential role of the oxidant-activated transcription factor aryl hydrocarbon receptor in the pathogenesis of AMD. Adv Exp Med Biol 723:51–59.
111. Mandai M, Kurimoto Y, Takahashi M 2017 Autologous induced stem-cell-derived retinal cells for macular degeneration. N Engl J Med 377:792–793.
112. Mares-Perlman JA, Brady WE, Klein R, VandenLangenberg GM, Klein BE, Palta M 1995 Dietary fat and age-related maculopathy. Arch Ophthalmol 113:743–748.
113. McHarg S, Clark SJ, Day AJ, Bishop PN 2015 Age-related macular degeneration and the role of the complement system. Mol Immunol 67:43–50.
114. McMahon M, Itoh K, Yamamoto M, Hayes JD 2003 Keap1-dependent proteasomal degradation of transcription factor Nrf2 contributes to the negative regulation of antioxidant response element-driven gene expression. J Biol Chem 278:21592–21600.
115. Meyers SM 1994 A twin study on age-related macular degeneration. Trans Am Ophthalmol Soc 92:775–843.
116. Milicevic N, Mazzaro N, de Bruin I, Wils E, Ten Brink J, Asbroek AT, Mendoza J, Bergen A, Felder-Schmittbuhl MP 2019 Rev-erbalpha and photoreceptor outer segments modulate the circadian clock in retinal pigment epithelial cells. Sci Rep 9:11790.
117. Mitter SK, Song C, Qi X, Mao H, Rao H, Akin D, Lewin A, Grant M, Dunn W Jr, Ding J, Bowes Rickman C, Boulton M 2014 Dysregulated autophagy in the RPE is associated with increased susceptibility to oxidative stress and AMD. Autophagy 10:1989–2005.
118. Mollema NJ, Yuan Y, Jelcick AS, Sachs AJ, von Alpen D, Schorderet D, Escher P, Haider NB 2011 Nuclear receptor Rev-erb alpha (Nr1d1) functions in concert with Nr2e3 to regulate transcriptional networks in the retina. PLoS One 6:e17494.
119. Mrowicka M, Mrowicki J, Szaflik JP, Szaflik M, Ulinska M, Szaflik J, Majsterek I 2017 Analysis of antioxidative factors related to AMD risk development in the polish patients. Acta Ophthalmol 95:530–536.
120. Murad N, Kokkinaki M, Gunawardena N, Gunawan MS, Hathout Y, Janczura KJ, Theos AC, Golestaneh N 2014 miR-184 regulates ezrin, LAMP-1 expression, affects phagocytosis in human retinal pigment epithelium and is downregulated in age-related macular degeneration. FEBS J 281:5251–5264.
121. Neale BM, Fagerness J, Reynolds R, Sobrin L, Parker M, Raychaudhuri S, Tan PL, Oh EC, Merriam JE, Souied E, Bernstein PS, Li B, Frederick JM, Zhang K, Brantley MA Jr, Lee AY, Zack DJ, Campochiaro B, Campochiaro P, Ripke S, et al. 2010 Genome-wide association study of advanced age-related macular degeneration identifies a role of the hepatic lipase gene (LIPC). Proc Natl Acad Sci U S A 107:7395–7400.
122. Newsome DA, Swartz M, Leone NC, Elston RC, Miller E 1988 Oral zinc in macular degeneration. Arch Ophthalmol 106:192–198.
123. Ni Dhubhghaill SS, Cahill MT, Campbell M, Cassidy L, Humphries MM, Humphries P 2010 The pathophysiology of cigarette smoking and age-related macular degeneration. Adv Exp Med Biol 664:437–446.
124. Nordgaard CL, Berg KM, Kapphahn RJ, Reilly C, Feng X, Olsen TW, Ferrington DA 2006 Proteomics of the retinal pigment epithelium reveals altered protein expression at progressive stages of age-related macular degeneration. Invest Ophthalmol Vis Sci 47:815–822.
125. Pardee KI, Xu X, Reinking J, Schuetz A, Dong A, Liu S, Zhang R, Tiefenbach J, Lajoie G, Plotnikov AN, Botchkarev A, Krause HM, Edwards A 2009 The structural basis of gas-responsive transcription by the human nuclear hormone receptor REV-ERBbeta. PLoS Biol 7:e43.
126. Pennesi ME, Neuringer M, Courtney RJ 2012 Animal models of age related macular degeneration. Mol Aspects Med 33:487–509.
127. Raghuram S, Stayrook KR, Huang P, Rogers PM, Nosie AK, McClure DB, Burris LL, Khorasanizadeh S, Burris TP, Rastinejad F 2007 Identification of heme as the ligand for the orphan nuclear receptors REV-ERBalpha and REV-ERBbeta. Nat Struct Mol Biol 14:1207–1213.
128. Ragsdale SW, Yi L, Bender G, Gupta N, Kung Y, Yan L, Stich TA, Doukov T, Leichert L, Jenkins PM, Bianchetti CM, George SJ, Cramer SP, Britt RD, Jakob U, Martens JR, Phillips GN Jr, Drennan CL 2012 Redox, haem and CO in enzymatic catalysis and regulation. Biochem Soc Trans 40:501–507.
129. Rahman I, MacNee W 1996 Role of oxidants/antioxidants in smoking-induced lung diseases. Free Radic Biol Med 21:669–681.
130. Ramkumar HL, Zhang J, Chan CC 2010 Retinal ultrastructure of murine models of dry age-related macular degeneration (AMD). Prog Retin Eye Res 29:169–190.
131. Rangasamy T, Cho CY, Thimmulappa RK, Zhen L, Srisuma SS, Kensler TW, Yamamoto M, Petrache I, Tuder RM, Biswal S 2004 Genetic ablation of Nrf2 enhances susceptibility to cigarette smoke-induced emphysema in mice. J Clin Invest 114:1248–1259.
132. Rattner A, Nathans J 2006 Macular degeneration:recent advances and therapeutic opportunities. Nat Rev Neurosci 7:860–872.
133. Reisman SA, Yeager RL, Yamamoto M, Klaassen CD 2009 Increased Nrf2 activation in livers from Keap1-knockdown mice increases expression of cytoprotective genes that detoxify electrophiles more than those that detoxify reactive oxygen species. Toxicol Sci 108:35–47.
134. Restrepo NA, Spencer KL, Goodloe R, Garrett TA, Heiss G, Bůžková P, Jorgensen N, Jensen RA, Matise TC, Hindorff LA, Klein BE, Klein R, Wong TY, Cheng CY, Cornes BK, Tai ES, Ritchie MD, Haines JL, Crawford DC 2014 Genetic determinants of age-related macular degeneration in diverse populations from the PAGE study. Invest Ophthalmol Vis Sci 55:6839–6850.
135. Rowan S, Jiang S, Chang ML, Volkin J, Cassalman C, Smith KM, Streeter MD, Spiegel DA, Moreira-Neto C, Rabbani N, Thornalley PJ, Smith DE, Waheed NK, Taylor A 2020 A low glycemic diet protects disease-prone Nrf2-deficient mice against age-related macular degeneration. Free Radic Biol Med 150:75–86.
136. Rowan S, Jiang S, Francisco SG, Pomatto LCD, Ma Z, Jiao X, Campos MM, Aryal S, Patel SD, Mahaling B, Riazuddin SA, Duh EJ, Lachke SA, Hejtmancik JF, de Cabo R, FitzGerald PG, Taylor A 2021 Aged Nrf2-null mice develop all major types of age-related cataracts. Invest Ophthalmol Vis Sci 62:10.
137. Sachdeva MM, Cano M, Handa JT 2014 Nrf2 signaling is impaired in the aging RPE given an oxidative insult. Exp Eye Res 119:111–114.
138. Saint-Geniez M, Kurihara T, Sekiyama E, Maldonado AE, D'Amore PA 2009 An essential role for RPE-derived soluble VEGF in the maintenance of the choriocapillaris. Proc Natl Acad Sci U S A 106:18751–18756.
139. SanGiovanni JP, Chew EY, Clemons TE, Ferris FL 3rd, Gensler G, Lindblad AS, Milton RC, Seddon JM, Sperduto RD 2007 The relationship of dietary carotenoid and vitamin A, E, and C intake with age-related macular degeneration in a case-control study:AREDS Report No. 22. Arch Ophthalmol 125:1225–1232.
140. SanGiovanni JP, Arking DE, Iyengar SK, Elashoff M, Clemons TE, Reed GF, Henning AK, Sivakumaran TA, Xu X, DeWan A, Agrón E, Rochtchina E, Sue CM, Wang JJ, Mitchell P, Hoh J, Francis PJ, Klein ML, Chew EY, Chakravarti A 2009 Mitochondrial DNA variants of respiratory complex I that uniquely characterize haplogroup T2 are associated with increased risk of age-related macular degeneration. PLoS One 4:e5508.
141. Schachat AP, Hyman L, Leske MC, Connell AM, Wu SY 1995 Features of age-related macular degeneration in a black population. The Barbados Eye Study Group. Arch Ophthalmol 113:728–735.
142. Seddon JM, Ajani UA, Mitchell BD 1997 Familial aggregation of age-related maculopathy. Am J Ophthalmol 123:199–206.
143. Seddon JM, Rosner B, Sperduto RD, Yannuzzi L, Haller JA, Blair NP, Willett W 2001 Dietary fat and risk for advanced age-related macular degeneration. Arch Ophthalmol 119:1191–1199.
144. Seddon JM, Cote J, Rosner B 2003 Progression of age-related macular degeneration:association with dietary fat, transunsaturated fat, nuts, and fish intake. Arch Ophthalmol 121:1728–1737.
145. Seddon JM, Reynolds R, Shah HR, Rosner B 2011 Smoking, dietary betaine, methionine, and vitamin D in monozygotic twins with discordant macular degeneration:epigenetic implications. Ophthalmology 118:1386–1394.
146. Seddon JM, Silver RE, Rosner B 2016 Response to AREDS supplements according to genetic factors:survival analysis approach using the eye as the unit of analysis. Br J Ophthalmol 100:1731–1737.
147. Shivarudrappa AH, Ponesakki G 2020 Lutein reverses hyperglycemia-mediated blockage of Nrf2 translocation by modulating the activation of intracellular protein kinases in retinal pigment epithelial (ARPE-19) cells. J Cell Commun Signal 14:207–221.
148. Sitaula S, Billon C, Kamenecka TM, Solt LA, Burris TP 2015 Suppression of atherosclerosis by synthetic REV-ERB agonist. Biochem Biophys Res Commun 460:566–571.
149. Smith CJ, Hansch C 2000 The relative toxicity of compounds in mainstream cigarette smoke condensate. Food Chem Toxicol 38:637–646.
150. Smith W, Mitchell P 1998 Family history and age-related maculopathy:the Blue Mountains Eye Study. Aust N Z J Ophthalmol 26:203–206.
151. Sohal RS, Orr WC 2012 The redox stress hypothesis of aging. Free Radical Biology and Medicine 52:539–555.
152. Solt LA, Wang Y, Banerjee S, Hughes T, Kojetin DJ, Lundasen T, Shin Y, Liu J, Cameron MD, Noel R, Yoo SH, Takahashi JS, Butler AA, Kamenecka TM, Burris TP 2012 Regulation of circadian behaviour and metabolism by synthetic REV-ERB agonists. Nature 485:62–68.
153. Souied EH, Benlian P, Amouyel P, Feingold J, Lagarde JP, Munnich A, Kaplan J, Coscas G, Soubrane G 1998 The epsilon4 allele of the apolipoprotein E gene as a potential protective factor for exudative age-related macular degeneration. Am J Ophthalmol 125:353–359.
154. Spencer KL, Hauser MA, Olson LM, Schmidt S, Scott WK, Gallins P, Agarwal A, Postel EA, Pericak-Vance MA, Haines JL 2008 Deletion of CFHR3 and CFHR1 genes in age-related macular degeneration. Hum Mol Genet 17:971–977.
155. Spraul CW, Lang GE, Grossniklaus HE, Lang GK 1999 Histologic and morphometric analysis of the choroid, Bruch's membrane, and retinal pigment epithelium in postmortem eyes with age-related macular degeneration and histologic examination of surgically excised choroidal neovascular membranes. Surv Ophthalmol 44 Suppl 1:S10–32.
156. Strauss O 2005 The retinal pigment epithelium in visual function. Physiol Rev 85:845–881.
157. Strunnikova N, Zhang C, Teichberg D, Cousins SW, Baffi J, Becker KG, Csaky KG 2004 Survival of retinal pigment epithelium after exposure to prolonged oxidative injury:a detailed gene expression and cellular analysis. Invest Ophthalmol Vis Sci 45:3767–3777.
158. Suh JH, Shenvi SV, Dixon BM, Liu H, Jaiswal AK, Liu RM, Hagen TM 2004 Decline in transcriptional activity of Nrf2 causes age-related loss of glutathione synthesis, which is reversible with lipoic acid. Proc Natl Acad Sci U S A 101:3381–3386.
159. Sulli G, Rommel A, Wang X, Kolar MJ, Puca F, Saghatelian A, Plikus MV, Verma IM, Panda S 2018 Pharmacological activation of REV-ERBs is lethal in cancer and oncogene-induced senescence. Nature 553:351–355.
160. Suzuki M, Betsuyaku T, Ito Y, Nagai K, Nasuhara Y, Kaga K, Kondo S, Nishimura M 2008 Down-regulated NF-E2-related factor 2 in pulmonary macrophages of aged smokers and patients with chronic obstructive pulmonary disease. Am J Respir Cell Mol Biol 39:673–682.
161. Tang Z, Ju Y, Dai X, Ni N, Liu Y, Zhang D, Gao H, Sun H, Zhang J, Gu P 2021 HO-1-mediated ferroptosis as a target for protection against retinal pigment epithelium degeneration. Redox Biol 43:101971.
162. Tarallo V, Hirano Y, Gelfand BD, Dridi S, Kerur N, Kim Y, Cho WG, Kaneko H, Fowler BJ, Bogdanovich S, Albuquerque RJ, Hauswirth WW, Chiodo VA, Kugel JF, Goodrich JA, Ponicsan SL, Chaudhuri G, Murphy MP, Dunaief JL, Ambati BK, et al. 2012 DICER1 loss and Alu RNA induce age-related macular degeneration via the NLRP3 inflammasome and MyD88. Cell 149:847–859.
163. Tong Y, Liao J, Zhang Y, Zhou J, Zhang H, Mao M 2010 LOC3≃5/HTRA1 gene polymorphisms and susceptibility to age-related macular degeneration:a HuGE review and meta-analysis. Mol Vis 16:1958–1981.
164. Tong Y, Zhang Z, Wang S 2022 Role of mitochondria in retinal pigment epithelial aging and degeneration. Front Aging 3:926627.
165. Urbańska K, Stępień PW, Nowakowska KN, Stefaniak M, Osial N, Chorągiewicz T, Toro MD, Nowomiejska K, Rejdak R 2022 The role of dysregulated miRNAs in the pathogenesis, diagnosis and treatment of age-related macular degeneration. Int J Mol Sci 23:7761.
166. van Asten F, Chiu CY, Agrón E, Clemons TE, Ratnapriya R, Swaroop A, Klein ML, Fan R, Chew EY 2019 No CFH or ARMS2 interaction with Omega-3 fatty acids, low versus high zinc, or β-carotene versus lutein and zeaxanthin on progression of age-related macular degeneration in the Age-Related Eye Disease Study 2:Age-Related Eye Disease Study 2 Report No. 18. Ophthalmology 126:1541–1548.
167. Varma R, Fraser-Bell S, Tan S, Klein R, Azen SP 2004 Prevalence of age-related macular degeneration in Latinos:the Los Angeles Latino eye study. Ophthalmology 111:1288–1297.
168. Vavvas DG, Small KW, Awh CC, Zanke BW, Tibshirani RJ, Kustra R 2018 CFH and ARMS2 genetic risk determines progression to neovascular age-related macular degeneration after antioxidant and zinc supplementation. Proc Natl Acad Sci U S A 115:E696–E704.
169. Vingerling JR, Dielemans I, Hofman A, Grobbee DE, Hijmering M, Kramer CF, de Jong PT 1995 The prevalence of age-related maculopathy in the Rotterdam Study. Ophthalmology 102:205–210.
170. Volonte D, Liu Z, Musille PM, Stoppani E, Wakabayashi N, Di YP, Lisanti MP, Kensler TW, Galbiati F 2013 Inhibition of nuclear factor-erythroid 2-related factor (Nrf2) by caveolin-1 promotes stress-induced premature senescence. Mol Biol Cell 24:1852–1862.
171. Wang AL, Lukas TJ, Yuan M, Du N, Tso MO, Neufeld AH 2009 Autophagy and exosomes in the aged retinal pigment epithelium:possible relevance to drusen formation and age-related macular degeneration. PLoS One 4:e4160.
172. Wang K, Zheng M, Lester KL, Han Z 2019 Light-induced Nrf2(-/-) mice as atrophic age-related macular degeneration model and treatment with nanoceria laden injectable hydrogel. Sci Rep 9:14573.
173. Wang L, Kondo N, Cano M, Ebrahimi K, Yoshida T, Barnett BP, Biswal S, Handa JT 2014 Nrf2 signaling modulates cigarette smoke-induced complement activation in retinal pigmented epithelial cells. Free Radic Biol Med 70:155–166.
174. Weismann D, Hartvigsen K, Lauer N, Bennett KL, Scholl HP, Charbel Issa P, Cano M, Brandstatter H, Tsimikas S, Skerka C, Superti-Furga G, Handa JT, Zipfel PF, Witztum JL, Binder CJ 2011 Complement factor H binds malondialdehyde epitopes and protects from oxidative stress. Nature 478:76–81.
175. Woldt E, Sebti Y, Solt LA, Duhem C, Lancel S, Eeckhoute J, Hesselink MK, Paquet C, Delhaye S, Shin Y, Kamenecka TM, Schaart G, Lefebvre P, Neviere R, Burris TP, Schrauwen P, Staels B, Duez H 2013 Rev-erb-alpha modulates skeletal muscle oxidative capacity by regulating mitochondrial biogenesis and autophagy. Nat Med 19:1039–1046.
176. Wong WL, Su X, Li X, Cheung CM, Klein R, Cheng CY, Wong TY 2014 Global prevalence of age-related macular degeneration and disease burden projection for 2020 and 2040:a systematic review and meta-analysis. Lancet Glob Health 2:e106–116.
177. Wu DM, Ji X, Ivanchenko MV, Chung M, Piper M, Rana P, Wang SK, Xue Y, West E, Zhao SR, Xu H, Cicconet M, Xiong W, Cepko CL 2021 Nrf2 overexpression rescues the RPE in mouse models of retinitis pigmentosa. JCI Insight 6:e145029.
178. Wu Y, Tian L, Huang Y 2015 Correlation between the interactions of ABCA4 polymorphisms and smoking with the susceptibility to age-related macular degeneration. Int J Clin Exp Pathol 8:7403–7408.
179. Xie T, Cai J, Yao Y, Sun C, Yang Q, Wu M, Xu Z, Sun X, Wang X 2021 LXA4 protects against blue-light induced retinal degeneration in human A2E-laden RPE cells and Balb-c mice. Ann Transl Med 9:1249.
180. Xiong W, MacColl Garfinkel AE, Li Y, Benowitz LI, Cepko CL 2015 NRF2 promotes neuronal survival in neurodegeneration and acute nerve damage. J Clin Invest 125:1433–1445.
181. Yang YP, Hsiao YJ, Chang KJ, Foustine S, Ko YL, Tsai YC, Tai HY, Ko YC, Chiou SH, Lin TC, Chen SJ, Chien Y, Hwang DK 2022 Pluripotent stem cells in clinical cell transplantation:focusing on induced pluripotent stem cell-derived RPE cell therapy in age-related macular degeneration. Int J Mol Sci 23:13794.
182. Yang Z, Camp NJ, Sun H, Tong Z, Gibbs D, Cameron DJ, Chen H, Zhao Y, Pearson E, Li X, Chien J, Dewan A, Harmon J, Bernstein PS, Shridhar V, Zabriskie NA, Hoh J, Howes K, Zhang K 2006 A variant of the HTRA1 gene increases susceptibility to age-related macular degeneration. Science 314:992–993.
183. Yang Z, Stratton C, Francis PJ, Kleinman ME, Tan PL, Gibbs D, Tong Z, Chen H, Constantine R, Yang X, Chen Y, Zeng J, Davey L, Ma X, Hau VS, Wang C, Harmon J, Buehler J, Pearson E, Patel S, et al. 2008 Toll-like receptor 3 and geographic atrophy in age-related macular degeneration. N Engl J Med 359:1456–1463.
184. You L, Peng H, Liu J, Cai M, Wu H, Zhang Z, Bai J, Yao Y, Dong X, Yin X, Ni J 2021 Catalpol protects ARPE-19 cells against oxidative stress via activation of the Keap1/Nrf2/ARE pathway. Cells 10:2635.
185. Young RW, Bok D 1969 Participation of the retinal pigment epithelium in the rod outer segment renewal process. J Cell Biol 42:392–403.
186. Zareparsi S, Reddick AC, Branham KE, Moore KB, Jessup L, Thoms S, Smith-Wheelock M, Yashar BM, Swaroop A 2004 Association of apolipoprotein E alleles with susceptibility to age-related macular degeneration in a large cohort from a single center. Invest Ophthalmol Vis Sci 45:1306–1310.
187. Zareparsi S, Buraczynska M, Branham KE, Shah S, Eng D, Li M, Pawar H, Yashar BM, Moroi SE, Lichter PR, Petty HR, Richards JE, Abecasis GR, Elner VM, Swaroop A 2005 Toll-like receptor 4 variant D299G is associated with susceptibility to age-related macular degeneration. Hum Mol Genet 14:1449–1455.
188. Zhang H, Davies KJA, Forman HJ 2015a Oxidative stress response and Nrf2 signaling in aging. Free Radic Biol Med 88:314–336.
189. Zhang J, Mitsuhashi T, Matsuo T, Yorifuji T, Hamada J, Liu Y 2021 Alcohol consumption and age-related macular degeneration:a systematic review and dose-response meta-analysis. Curr Eye Res 46:1900–1907.
190. Zhang Y, Fang B, Emmett MJ, Damle M, Sun Z, Feng D, Armour SM, Remsberg JR, Jager J, Soccio RE, Steger DJ, Lazar MA 2015b GENE REGULATION. Discrete functions of nuclear receptor Rev-erbalpha couple metabolism to the clock. Science 348:1488–1492.
191. Zhang Y, Wong WT 2021 Innate immunity in age-related macular degeneration. Adv Exp Med Biol 1256:121–141.
192. Zhao X, Gao M, Liang J, Chen Y, Wang Y, Wang Y, Xiao Y, Zhao Z, Wan X, Jiang M, Luo X, Wang F, Sun X 2021 SLC7A11 reduces laser-induced choroidal neovascularization by inhibiting RPE ferroptosis and VEGF production. Front Cell Dev Biol 9:639851.
193. Zhao Z, Chen Y, Wang J, Sternberg P, Freeman ML, Grossniklaus HE, Cai J 2011 Age-related retinopathy in NRF2-deficient mice. PLoS One 6:e19456.
194. Zorov DB, Juhaszova M, Sollott SJ 2014 Mitochondrial reactive oxygen species (ROS) and ROS-induced ROS release. Physiol Rev 94:909–950.

age-related macular degeneration; antioxidant; nuclear factor erythroid-2-related factor 2; oxidative stress; retinal pigment epithelium; REV-ERBα

Copyright: © 2023 Neural Regeneration Research