Substantial epidemiological evidence suggests that chronic inflammation is associated with various aging phenotypes, such as metabolic homeostasis, immune senescence, and changes in body composition (Franceschi and Campisi, 2014). Inflammation is also implicated in the etiology of multiple aging-related diseases (Sanada et al., 2018), including cardiovascular disease and cancer (Gan et al., 2004; Pawelec et al., 2014; Singh et al., 2019). Dietary and lifestyle exposures likely contribute to higher chronic systemic inflammation (Cavicchia et al., 2009; Barbaresko et al., 2013; Kantor et al., 2013), and genetic variation may too (Shen and Ordovas, 2009). Since reactive oxygen species (ROS) play an important role in inflammation and the pathogenesis of many inflammatory diseases (Mittal et al., 2014), and antioxidant enzymes (AE) and DNA base excision repair (BER) mechanisms contribute to protecting cells from ROS-induced deleterious effects (Sun, 1990; Seeberg et al., 1995; Mates, 2000), polymorphisms in AE and BER genes may be especially important in regulating systemic inflammation.

Substantial basic science and human study evidence supports that multiple dietary constituents and lifestyle factors contribute to systemic inflammation, and recent epidemiologic evidence consistently, strongly supports that dietary and lifestyle factors collectively are particularly strongly associated with circulating concentrations of inflammation biomarkers and chronic diseases for which inflammation is considered an important risk factor. Circulating concentrations of high sensitivity C-reactive protein (hsCRP), an acute phase protein produced in the liver and released into the circulation, has been used extensively clinically and in epidemiologic studies (Byrd et al., 2019) as a biomarker of inflammation. Although, for some applications, it has some limitations (e.g., increases in response to infection and other inflammatory medical conditions (Sproston and Ashworth, 2018)), it is reliably measured (Ridker, 2004), is the most widely used biomarker of inflammation in epidemiologic studies, and is highly correlated with other biomarkers of inflammation (Byrd et al., 2019). We recently reported the development and validation of novel, questionnaire-based, dietary and lifestyle inflammation scores, the DIS and LIS, respectively (Byrd et al., 2019). The components of the predominantly whole foods-based, 19-component DIS and the 4-component LIS [comprising physical activity, body mass index (BMI), alcohol intake, and smoking status] are weighted according to the strengths of their associations with a panel of inflammation biomarkers (including hsCRP) in a diverse population of Black and White male and female adults from the 48 contiguous United States. The DIS was more strongly associated with inflammation biomarkers than were other reported dietary inflammation indices in three populations (one of which included participants in the present study), and the strongest associations were among those who were jointly in the highest DIS and LIS categories (Byrd et al., 2019).

Just as individual dietary and lifestyle factors, for the most part, tend to be modestly and inconsistently associated with inflammation, but collectively are strongly and consistently associated with it (Byrd et al., 2019), it is unlikely that single, relatively common AE or BER polymorphisms would be strongly associated with systemic inflammation, but collectively they might. Genetic risk scores (GRS) that combine multiple variants in multiple genes were previously reported and found to be associated with chronic disease outcomes (Wang et al., 2017; Chalmer et al., 2018; Korologou-Linden et al., 2019a; Korologou-Linden et al., 2019b; Leonenko et al., 2019; Mavaddat et al., 2019; Watt et al., 2019; Zheutlin et al., 2019; Li et al., 2020; Mosley et al., 2020; Sipeky et al., 2020). Associations of AE and BER GRS with colorectal adenoma were previously reported (Wang et al., 2017); however, there are no reported associations of these or other GRS, alone or in interaction with dietary and lifestyle exposures, with inflammation biomarkers.

Accordingly, herein we report the development of inflammation-specific AE and BER GRS and their associations with circulating hsCRP concentrations, individually and jointly with the DIS and LIS in two pooled cross-sectional studies.

Selected characteristics of the participants according to hsCRP concentration tertiles are summarized in Table 1. Plasma hsCRP concentrations ranged from 0.14 to 32.67 μg/ml. Participants in the highest relative to the lowest hsCRP tertile were more likely to be female, a current smoker, and, if a woman, to take HRT; less likely to be formally educated and to have a heavy alcohol intake; and, on average, had a higher BMI, DIS, and LIS, and lower total vitamin C intake.

Mean plasma hsCRP concentrations according to AE and BER GRS categories are shown in Table 2. There was a dose-response pattern for increasing mean hsCRP concentrations across the BER GRS tertiles, and the mean hsCRP concentration among those in the highest relative to the lowest BER GRS tertile was, proportionately, statistically significantly 57.4% higher. Among those in the high relative to the low AE GRS category, the mean hsCRP concentration was estimated to be 13.9% higher, but the finding was not statistically significant. Adjustment for multiple covariates (e.g., sex, the DIS and the LIS) negligibly affected the estimates.

The mean plasma hsCRP concentrations in the joint dichotomized AE/BER GRS categories are shown in Table 3. The lowest joint AE/BER GRS category was considered the reference group (the hypothesized lowest risk group). Although we found no statistically significant interaction between the AE and BER GRS, the mean hsCRP concentration was highest (62.7% higher) among those in the highest relative to the lowest joint AE/BER GRS category (p = 0.006). Adjustment for multiple covariates negligibly affected the estimates.

We previously reported that in the larger (n = 423) pooled MAP studies population, which did not exclude participants missing genotyping data or who were not White, among those in the highest relative to the lowest DIS and LIS quartiles, multivariable-adjusted mean circulating hsCRP concentrations were 31.6% (p = 0.02) and 129% higher (p < 0.001), respectively (Byrd et al., 2020). In the present, more restricted pooled MAP studies population, among those in the highest relative to the lowest DIS and LIS tertiles, multivariable-adjusted mean circulating hsCRP concentrations were 18.5% (p = 0.66) and 120% higher (p < 0.001), respectively (Supplementary Table S8).

Multivariable-adjusted mean plasma hsCRP concentrations across DIS and LIS tertiles, stratified by dichotomized BER and AE GRS are shown in Table 4. We observed no strong or statistically significant evidence for effect modification of the DIS- and LIS-hsCRP associations by either GRS. However, there were some suggestions that the positive DIS-hsCRP association was slightly stronger among those with a high AE GRS, and that the positive LIS-hsCRP association was slightly stronger among those with a low AE GRS. Specifically, among those with a high AE GRS, the mean hsCRP concentration among those in the highest relative to the lowest DIS tertile was estimated to be 45.5% higher, whereas among those with a low AE GRS it was estimated to be 13.3% higher. Among those with a low AE GRS, the mean hsCRP concentration among those in the highest relative to the lowest LIS tertile was 194.3% higher, whereas among those with a high AE GRS it was estimated to be 68.6% higher.

Our findings suggest that certain BER genotypes (MUTYH, SMUG1, TDG, UNG, and XRCC1) collectively may be associated with systemic inflammation, as reflected by circulating hsCRP concentrations. We found no definitive evidence that AE genotypes collectively are associated with systemic inflammation; however, plasma hsCRP concentrations were estimated to be modestly higher among individuals with more AE risk variants (i.e., had a higher AE GRS). We also found suggestions that the AE and BER GRS may i) modestly modify the associations of collective dietary and lifestyle exposures with systemic inflammation, and ii) interact with each other such that those with a higher relative to a lower joint BER/AE GRS may have modestly higher systemic inflammation. These findings from this preliminary investigation support further investigations in larger, general populations.

Previous basic science studies, reviewed extensively elsewhere (Sun, 1990; Seeberg et al., 1995; Mates, 2000; Di Virgilio, 2004; Kidane et al., 2014; Yang and Lee, 2015; Li and Chen, 2018), provide biological plausibility for investigating possible associations of AE and BER genotypes with inflammation. Briefly, higher levels of reactive oxygen and nitrogen species (RONS), from both exogenous and endogenous sources, damage cell structures and DNA, eliciting inflammatory responses (which in turn can increase RONS levels) and DNA BER, respectively. Antioxidant enzymes lower RONS levels, or neutralize their excessive cellular oxidation effects (Yang and Lee, 2015). As examples, of the genes included in our AE GRS, CAT defends against superoxide and hydrogen peroxide and constitutes a primary defense against oxidative stress (Röhrdanz and Kahl, 1998), and MnSoD prevents disruption of mitochondrial membrane potential and catalyzes dismutation of superoxide radicals (Röhrdanz and Kahl, 1998; Mäntymaa et al., 2000). As reviewed elsewhere (Kidane et al., 2014; Li and Chen, 2018), there is increasing evidence for interplay between DNA repair and inflammation. DNA damage is present during inflammation, accumulates during chronic inflammation, and DNA repair plays an important role in counteracting this damage (Kidane et al., 2014). RONS-induced DNA damage is generally repaired by the BER pathway (Wallace et al., 2012), and mutations in BER genes are associated with chronic inflammation. The DNA damage response (DDR) directly activates several transcription factors, such as NF-κB and interferon regulatory factors (IRFs) (McCool and Miyamoto, 2012). These transcription factors induce the expression of various immune genes, including those for inflammatory cytokines and chemokines. In addition, the DDR and oxidative stress induce the expression of several ligands for activating immune receptors. Although the exact mechanisms for these observations are not fully elucidated, possible mechanisms could include that accumulation of RONS-induced lesions (or associated mutations) in, for example, the mitochondrial genome, could lead to respiratory defects that increase intracellular ROS concentrations (Li and Chen, 2018); and oxidative DNA damage excision fragments could activate sensing programs such as cGAS/STING (Li and Chen, 2018), which may promote inflammatory outcomes. Evidence such as outlined above supports that DNA BER may play a role in controlling RONS levels, and that the efficiency of such control may affect RONS-induced inflammation. As examples of the polymorphic BER-related genes included in our BER GRS, MUTYH contributes to the repair of one of the most frequent and stable forms of oxidative damage by removing the mismatched 8-oxoG adenine (Sampson et al., 2005; Nielsen et al., 2011); SMUG1 removes uracil from single- and double-stranded DNA in nuclear chromatin (Nilsen et al., 2001; Broderick et al., 2006); TDG has been implicated in DNA demethylation and could repair G/T and G/U mismatches via removing thymine and uracil moieties (He et al., 2011; Wu and Zhang, 2017); UNG repairs mutagenic G/U mismatches caused by deamination of cytosine (Krokan et al., 2001); and XRCC1 plays a role in repairing single-stranded DNA breaks (Duell et al., 2000; Thompson and West, 2000). In summary, AE and BER genes reduce systemic inflammation via reducing RONS production, neutralizing oxidation, and repairing oxidative damage in humans.

Recently, GRS, also called polygenic risk scores (PRS), have been used as tools for investigating risk for various diseases (Wang et al., 2017; Chalmer et al., 2018; Korologou-Linden et al., 2019a; Korologou-Linden et al., 2019b; Leonenko et al., 2019; Mavaddat et al., 2019; Watt et al., 2019; Zheutlin et al., 2019; Li et al., 2020; Mosley et al., 2020; Sipeky et al., 2020), and support the concept of investigating associations of GRS with various outcomes. However, our study is the first to report associations of GRS with an inflammation biomarker. In epidemiologic studies, various GRS/PRS were reported to be associated with multiple diseases, including colorectal cancer (Wang et al., 2017), prostate cancer (Sipeky et al., 2020), breast cancer (Mavaddat et al., 2019; Watt et al., 2019), Alzheimer’s disease (Korologou-Linden et al., 2019a; Korologou-Linden et al., 2019b; Leonenko et al., 2019), and psychiatric diseases (Chalmer et al., 2018; Zheutlin et al., 2019). To our knowledge, there are no reported investigations of an AE GRS, and only one (Wang et al., 2017) of a BER GRS. That analysis used data pooled from three colonoscopy-based case-control studies (408 adenoma cases and 604 controls), and constructed a BER GRS based on 65 individual SNPs in 15 BER genes (Wang et al., 2017). That study’s findings suggested that colorectal adenoma risk among participants in the highest relative to the lowest tertile of that BER GRS was statistically significantly higher (odds ratio = 2.07, 95% CI = 1.26–3.40) (Wang et al., 2017). A cross-sectional study of hemodialysis patients (n = 167) investigated associations of BER genetic polymorphisms (3 SNPs; 2 for MUTYH and 1 for OGG1, but not the same as those in our study) with inflammation biomarkers (Cai et al., 2012). The findings suggested that two variant MUTYH genotypes, singly and in combination with an OGG1 variant, were associated with higher IL-1β and IL-6 concentrations in that patient population (Cai et al., 2012). However, as noted, the authors investigated only three SNPs from BER genes and a GRS was not constructed (Cai et al., 2012).

Several studies investigated BER genetic variants as potential effect modifiers of associations of dietary and lifestyle exposures with colorectal adenoma risk (Corral et al., 2013; Wang et al., 2017), and support the concept of GRS as potential modifiers of associations of environmental exposures with various outcomes. However, our study is the first to report such potential effect modification in relation to an inflammation biomarker. Results from the three pooled case-control studies noted above suggested that the positive association of a higher balance of pro-over anti-oxidant dietary and lifestyle exposures with colorectal adenoma risk was stronger among those with more relative to fewer BER genetic variants (Wang et al., 2017). In a matched case-control study (677 adenoma cases and 691 controls), BER gene SNPs (although the SNPs investigated were not the same as those in our study) were estimated to be potential effect modifiers of the associations of colorectal adenoma with smoking (MUTYH P _interaction = 0.002, OGG1 P _interaction = 0.01, and FEN1 P _interaction = 0.01), alcohol (LIG3 P _interaction = 0.02), and dietary folate (LIG3 P _interaction = 0.02) (Corral et al., 2013).

Our study has several limitations. First, the GRS were calculated based on tagSNPs in only AE and BER genes in the present, relatively small study population, rather than via a genome-wide association study in a large population, and involved multiple comparisons. However, to our knowledge, our study is the first to report associations of GRS with an inflammation biomarker and provides support and preliminary data for future more definitive investigations. Second, our study population included only White participants who went for outpatient colonoscopy, which may limit the generalizability of our findings. Third, although our tagSNP approach comprehensively covered all common SNP variation in AE and BER genes, it was not possible to cover rare variation (MAF ≤5%); thus, potential influential SNPs may have been excluded. Fourth, we had measurements on only one biomarker of inflammation, hsCRP. A panel of biomarkers to represent multiple aspects of inflammation may more accurately represent systemic inflammation (Byrd et al., 2019), and as noted in Section 1, hsCRP has certain limitations. However, our study population was in general good health; we excluded those with inflammatory bowel disease, cancer, familial adenomatous polyposis, or extreme hsCRP values; assessed the use of NSAIDs as potential confounders and effect modifiers; and hsCRP measurements in such a population are highly correlated with those of other inflammation biomarkers (Byrd et al., 2019). Other limitations include the general limitations of using FFQs to assess diet, such as recall error, limited food choices, and issues regarding capturing seasonal intake patterns. However, we used a previously validated FFQ (Willett et al., 1985), and in our study, recall error would be expected to be non-differential (participants reported dietary and lifestyle exposures before hsCRP measurements), which would be expected to attenuate the DIS and LIS results.

Strengths of our study include the collection and assessment of extensive dietary, lifestyle, and medical data as potential confounding factors, the high quality of the laboratory measurements, and the inclusion of both men and women. Also, to our knowledge, our study is the first to report i) associations of AE and BER GRS with a biomarker of inflammation in humans, and ii) AE and BER GRS as potential effect modifiers of the associations of collective dietary and lifestyle exposures with a biomarker of inflammation.

In conclusion, our findings, taken together with previous literature, suggest that, collectively, genotypes of DNA BER genes may be associated with systemic inflammation in humans. Our study serves as a pilot study that supports further investigations of inflammation-specific BER and AE GRS, individually and in interaction with each other and dietary and lifestyle inflammation scores, in larger, general populations.