Edward Loane

Transport and Retinal Capture of Lutein and Zeaxanthin with Reference to Age-related Macular Degeneration

Age-related macular degeneration (AMD) is the most common cause of irreversible blindness in the elderly population in the western world. The etiology and pathogenesis of this disease remain unclear. However, there is an increasing body of evidence supporting the hypothesis that the macular pigment carotenoids, lutein and zeaxanthin, play an important role in protection against AMD, by filtering out blue light at a pre-receptoral level, or by quenching free radicals. Lutein and zeaxanthin are dietary xanthophyll carotenoids, which are delivered to the retina via plasma lipoproteins. The biological mechanisms governing retinal capture and accumulation of lutein and zeaxanthin, to the exclusion of other carotenoids, are still poorly understood. Although these mechanisms remain unclear, it is possible that selective capture of these carotenoids is related to lipoprotein, or apolipoprotein, function and profile. Xanthophyll-binding proteins appear to play an important role in the retinal capture of the xanthophyll carotenoids. The Pi isoform of GSTP1 has been isolated as a specific binding protein for zeaxanthin. The binding protein responsible for retinal uptake of lutein remains elusive. This article reviews the literature germane to the mechanisms involved in the capture, accumulation and stabilization of lutein and zeaxanthin by the retina, and the processes involved in their transport in serum.

The rationale and evidence base for a protective role of macular pigment in age-related maculopathy

Age-related maculopathy (ARM) remains the most common cause of blind registration in people aged 50 years or over in the developed world, and its prevalence continues to rise. Although effective new treatments have become available in the recent past, these are expensive and cumbersome to the healthcare provider and to the patient, and many cases remain resistant to such therapy. There is a biologically plausible rationale whereby macular pigment, which is entirely of dietary origin, may prevent or delay the onset, or ameliorate the clinical course, of ARM. In this article, we review this rationale, and critically appraise the current evidence base germane to the use of supplements containing the macular carotenoids in patients with, or at risk of developing, ARM.

Augmentation of macular pigment following implantation of blue light-filtering intraocular lenses at the time of cataract surgery.

PURPOSE (Photo)-oxidative stress is believed to play a role in the pathogenesis of age-related macular degeneration (AMD), with the threshold for retinal damage being lowest for short-wavelength (blue) light. Macular pigment (MP), consisting of the carotenoids lutein (L), zeaxanthin (Z) and meso-Z, has a maximum absorption at 460 nm and protects the retina from (photo)-oxidative injury. This study was designed to investigate whether the blue light-filtering properties of the Alcon AcrySof Natural intraocular lens (ANIOL) implanted during cataract surgery affects MP optical density (MPOD). METHODS Forty-two patients scheduled for cataract surgery were recruited for the study. These patients all had a preoperative best corrected visual acuity rating (BCVAR) of at least 0.5 (logMAR) in the study eye. The patients were randomized to have either the standard Alcon AcrySof three-piece acrylic intraocular lens (AIOL) (controls) or the ANIOL implanted at the time of cataract surgery. The spatial profile of MPOD (i.e., at 0.25 degrees, 0.5 degrees, 1.0 degrees, and 1.75 degrees eccentricity) was measured with customized heterochromatic flicker photometry (cHFP) 1 week before and 1 week after surgery, and at 3, 6, and 12 months after surgery. Serum concentrations of L and Z were also measured at each study visit. RESULTS There was a highly significant and positive correlation between all MPODs (e.g., at 0.25 degrees) recorded 1 week before and after surgery in eyes with an AIOL implant (r = 0.915, P < 0.01; paired samples t-test, P = 0.631) and in those ANIOL implants (r = 0.868, P < 0.01; paired samples t-test, P = 0.719). Average MPOD across the retina increased significantly with time (after 3 months) in the ANIOL group (repeated-measures, general linear model, P < 0.05), but remained stable in the AIOL group (repeated-measures, general linear model, P > 0.05). There were no significant time or lens effects observed for serum L over the study period (P > 0.05). There was a significant time effect for serum Z over the study period (P < 0.05), but not a significant time/lens interaction (P > 0.05). CONCLUSIONS Customized HFP can reliably measure the MPOD spatial profile in the presence of lens opacity, and cataract surgery does not artifactually alter MPOD readings. This study also provides evidence that implanting an IOL that filters blue light is associated with augmentation of MPOD in the absence of raised serum concentrations of L and Z. However, further and longitudinal study is needed to assess whether the observed increase in MPOD after implantation of blue-filtering IOLs is associated with reduced risk of AMD development and/or progression.

Foveal anatomic associations with the secondary peak and the slope of the macular pigment spatial profile.

PURPOSE To investigate the reproducibility of the macular pigment (MP) spatial profile by using heterochromatic flicker photometry (HFP) and to relate the MP spatial profile to foveal architecture. METHODS Sixteen healthy subjects (nine had the typical exponential MP spatial profile [group 1]; seven had a secondary peak MP spatial profile [group 2]) were recruited. The MP spatial profile was measured on three separate occasions. Six radiance measurements were obtained at each locus (0.25 degrees , 0.5 degrees , 1 degrees , and 1.75 degrees eccentricity; reference point, 7 degrees ). Foveal architecture was assessed by optical coherence tomography (OCT). RESULTS Subjects who had the typical decline profile, had this profile after averaging repeated measures (group 1). Subjects who had a secondary peak, displayed the secondary peak after repeated measures were averaged (group 2). Mean SD foveal width in group 1 was significantly narrower than mean SD foveal width in group 2 (1306 +/- 240 microm and 1915 +/- 161 microm, respectively; P < 0.01). This difference remained after adjustment for sex (P < 0.001). Foveal width was significantly related to mean foveal MP, with adjustment for sex (r = 0.588, P = 0.021). Foveal profile slope was significantly related to MP spatial profile slope, after removal of an outlier (r = 0.591, P = 0.020). CONCLUSIONS HFP reproducibly measures MP spatial profile. Secondary peaks seen in the MP spatial profile cannot be attributed to measurement error and are associated with wider foveas. The slope of an individuals MP spatial profile is related to foveal slope, with a steeper MP distribution associated with a steeper foveal depression.

Measurement of Macular Pigment Optical Density Using Two Different Heterochromatic Flicker Photometers

Purpose: To compare macular pigment optical density using two different heterochromatic flicker photometers. Methods: We measured macular pigment optical density in 121 healthy subjects using heterochromatic flicker photometry. Results: The mean (±SD) macular pigment optical density measured using the Maculometer was 0.394 (±0.170), and that using the Densitometer was 0.395 (±0.189). The difference in measurements on each instrument was influenced by age and macular pigment levels. Conclusions: On average, there is no difference in measurements provided by these two instruments. The Maculometer tends to underestimate macular pigment in older subjects and/or those with higher macular pigment compared with the Densitometer.

The respective relationships between lipoprotein profile, macular pigment optical density, and serum concentrations of lutein and zeaxanthin.

PURPOSE The dietary carotenoids lutein (L) and zeaxanthin (Z) are transported on lipoproteins in the serum. The mechanism of delivery of L and Z to the macula, where they constitute macular pigment (MP), is poorly understood. This study was an investigation of the respective relationships between serum lipoprotein profile, MP optical density (MPOD), and serum L and Z. METHODS Three hundred two healthy subjects were recruited; 211 (69.9%) were women. Demographic and health details were recorded. Fasting blood samples were taken for lipoprotein analysis by spectrophotometric assay and L and Z analysis by high-performance liquid chromatography. MPOD was measured by heterochromatic flicker photometry. RESULTS The mean ± SD (range) age of all subjects was 48 ± 11 (21-66) years. There was a statistically significant and positive association between serum L concentration and both serum cholesterol concentration (r = 0.239, P < 0.001) and serum HDL concentration (r = 0.324, P < 0.001), but not with serum LDL concentration (r = 0.095, P = 0.101). There was a statistically significant but inverse association between serum triglyceride concentration and total MPOD (r = -0.118, P = 0.044). There was no significant association between MPOD and serum cholesterol concentration or serum HDL concentration (P > 0.05). CONCLUSIONS The findings are consistent with the hypothesis that HDL is important for the transport of L in serum. The mechanism(s) whereby L and Z are captured by the macula and whether the serum (apo)lipoprotein profile is important in the transfer of the carotenoids from serum to retina merit further study.

Apolipoprotein E genotype is associated with macular pigment optical density.

PURPOSE Age-related macular degeneration (AMD) is the most common cause of blindness in older people in developed countries, and risk factors for this condition may be classified as genetic and environmental. Apolipoprotein E is putatively involved in the transport of the macular pigment (MP) carotenoids lutein (L) and zeaxanthin (Z) in serum and may also influence retinal capture of these compounds. This study was designed to investigate the relationship between macular pigment optical density (MPOD) and ApoE genotype. METHODS This was a cross-sectional study of 302 healthy adult subjects. Dietary intake of L and Z was assessed by food frequency questionnaire, and MPOD was measured by customized heterochromatic flicker photometry. Serum L and Z were measured by HPLC. ApoE genotyping was performed by direct polymerase chain reaction amplification and DNA nucleotide sequencing from peripheral blood. RESULTS Genotype data were available on 300 of the 302 (99.3%) subjects. The mean (+/- SD) age of the subjects in this study was 47.89 +/- 11.05 (range, 21-66) years. Subjects were classed into one of three ApoE genotype groups, as follows: group 1, epsilon2epsilon2 or epsilon2epsilon3; group 2, epsilon3epsilon3; group 3, epsilon2epsilon4 or epsilon3epsilon4 or epsilon4epsilon4. All three groups were statistically comparable in terms of age, sex, body mass index, cigarette smoking, and dietary and serum levels of L and Z. There was a statistically significant association between ApoE genotype and MPOD. Subjects who had at least one epsilon4 allele had a higher MPOD across the macula than subjects without this allele (group 1 MPOD area, 0.70 +/- 0.40; group 2 MPOD area, 0.67 +/- 0.42; group 3 MPOD area, 0.85 +/- 0.46; one-way ANOVA, P = 0.014. CONCLUSIONS These results suggest that ApoE genotype status is associated with MPOD. This association may explain, at least in part, the putative protective effect of the epsilon4 allele for AMD and is consistent with the view that apolipoprotein profile influences the transport and/or retinal capture of circulating L and/or Z.

Risk Factors for Age-Related Maculopathy

Age-related maculopathy (ARM) is the leading cause of blindness in the elderly. Although beneficial therapeutic strategies have recently begun to emerge, much remains unclear regarding the etiopathogenesis of this disorder. Epidemiologic studies have enhanced our understanding of ARM, but the data, often conflicting, has led to difficulties with drawing firm conclusions with respect to risk for this condition. As a consequence, we saw a need to assimilate the published findings with respect to risk factors for ARM, through a review of the literature appraising results from published cross-sectional studies, prospective cohort studies, case series, and case control studies investigating risk for this condition. Our review shows that, to date, and across a spectrum of epidemiologic study designs, only age, cigarette smoking, and family history of ARM have been consistently demonstrated to represent risk for this condition. In addition, genetic studies have recently implicated many genes in the pathogenesis of age-related maculopathy, including Complement Factor H, PLEKHA 1, and LOC387715/HTRA1, demonstrating that environmental and genetic factors are important for the development of ARM suggesting that gene-environment interaction plays an important role in the pathogenesis of this condition.

The association between macular pigment optical density and CFH, ARMS2, C2/BF, and C3 genotype

Age-related macular degeneration (AMD) is the most common cause of blindness in older people in developed countries, and risk for this condition may be classified as genetic or environmental, with an interaction between such factors predisposing to this disease. This study investigated the relationship between AMD risk genes, macular pigment optical density (MPOD), which may protect against AMD, and serum concentrations of the macular carotenoids, lutein (L) and zeaxanthin (Z). This was a cross-sectional study of 302 healthy adult subjects. Dietary intake of L and Z was assessed by food frequency questionnaire, and MPOD was measured by customized heterochromatic flicker photometry. We also calculated MPOD Area as the area of MP under the spatial profile curve, to reflect MP across the macula. Serum L and Z were measured by HPLC. Genotyping of tag SNPs in the genes CFH, ARMS2, C3, C2 and BF was undertaken with multiplex polymerase chain reaction (PCR) and primer extension methodology (ABI Snapshot, ABI Warrington UK) on DNA extracted from peripheral blood. The mean ± SD (range) age of the subjects in this study was 48 ± 11 (21-66) years. There was a statistically significant association between CFH genotype and family history of AMD, with subjects having two non-risk CFH haplotypes (n = 35), or one non-risk and one protective CFH haplotype (n = 33), being significantly more likely to have a negative family history of AMD (Pearson Chi square: p = 0.001). There was no significant association between the AMD risk genes investigated and either MPOD (One way ANOVA: p > 0.05) or serum concentrations of L or Z (One way ANOVA: p > 0.05, for both). Subjects who were homozygous for risk alleles of both CFH and ARMS2 (n = 4) had significantly lower MPOD at 0.5° and 1° retinal eccentricity (Independent samples t test: p < 0.05) and lower MPOD Area which approached statistical significance (Independent samples t test: p = 0.058), compared to other subjects (n = 291). In conclusion, this study did not detect an association between individual AMD risk genotypes and the putatively protective MP, or serum concentrations of its constituent carotenoids. However, the combination of homozygous risk alleles at both CFH and ARMS2 loci was associated with significantly lower MPOD centrally, despite comparable serum concentrations of the macular carotenoids. These findings suggest that the maculae of subjects at very high genetic risk of AMD represent a hostile environment for accumulation and/or stabilization of MP.

Investigation of Genetic Variation in Scavenger Receptor Class B, Member 1 (SCARB1) and Association with Serum Carotenoids

OBJECTIVE To investigate association of scavenger receptor class B, member 1 (SCARB1) genetic variants with serum carotenoid levels of lutein (L) and zeaxanthin (Z) and macular pigment optical density (MPOD). DESIGN A cross-sectional study of healthy adults aged 20 to 70. PARTICIPANTS We recruited 302 participants after local advertisement. METHODS We measured MPOD by customized heterochromatic flicker photometry. Fasting blood samples were taken for serum L and Z measurement by high-performance liquid chromatography and lipoprotein analysis by spectrophotometric assay. Forty-seven single nucleotide polymorphisms (SNPs) across SCARB1 were genotyped using Sequenom technology. Association analyses were performed using PLINK to compare allele and haplotype means, with adjustment for potential confounding and correction for multiple comparisons by permutation testing. Replication analysis was performed in the TwinsUK and Carotenoids in Age-Related Eye Disease Study (CAREDS) cohorts. MAIN OUTCOME MEASURES Odds ratios for MPOD area, serum L and Z concentrations associated with genetic variations in SCARB1 and interactions between SCARB1 and gender. RESULTS After multiple regression analysis with adjustment for age, body mass index, gender, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglycerides, smoking, and dietary L and Z levels, 5 SNPs were significantly associated with serum L concentration and 1 SNP with MPOD (P<0.01). Only the association between rs11057841 and serum L withstood correction for multiple comparisons by permutation testing (P<0.01) and replicated in the TwinsUK cohort (P = 0.014). Independent replication was also observed in the CAREDS cohort with rs10846744 (P = 2×10(-4)), an SNP in high linkage disequilibrium with rs11057841 (r(2) = 0.93). No interactions by gender were found. Haplotype analysis revealed no stronger association than obtained with single SNP analyses. CONCLUSIONS Our study has identified association between rs11057841 and serum L concentration (24% increase per T allele) in healthy subjects, independent of potential confounding factors. Our data supports further evaluation of the role for SCARB1 in the transport of macular pigment and the possible modulation of age-related macular degeneration risk through combating the effects of oxidative stress within the retina. FINANCIAL DISCLOSURE(S) Proprietary or commercial disclosures may be found after the references.