Association of vitamin D receptor TaqI and ApaI genetic polymorphisms with nephrolithiasis and end stage renal disease: a meta-analysis

In human skin, solar rays facilitate the formation of vitamin D3 from 7-dehydrocholesterol. The vitamin D3 undergoes two-step hydroxylation to form 25-hydroxy vitamin D3 (25-OHD3) and biologically active 1,25-dihydroxyvitamin D3 (1,25-(OH)₂D3) [1]. Vitamin D receptor (VDR) is a ligand-activated transcriptional factor requiring 1,25(OH)2D for its activation [2]. The deficiency of 25OHD or VDR is reported to activate renin-angiotensin system resulting in high angiotensin II levels, which damage renal parenchyma leading to increased risk for renal disease [3]. Considering the pivotal role of VDR in maintaining normal renal function, a number of studies have explored the possibility of association of VDR gene polymorphisms with renal disease risk. Among VDR polymorphisms reported to date, ApaI, and TaqI are widely studied for their association with ESRD, NL and DN [4‐6]. The ApaI variant (rs7975232), which results in A to C transition, is located in the intron 8 of VDR gene, while TaqI variant (rs731236), which results in T to C transition is located in exon 9 [7].

The rs7975232 (NG_008731.1:g.64978G > T) is an intronic variant predicted to influence splice site changes that might affect the translation of VDR. The frequency of this variant is high as evidenced by 734 and 16,751 homozygous mutants in 1000G and ExAC databases. The rs731236 (NG_008731.1:g.65058 T > C) variant is near the exon-intron boundary (GCTG/attg) and hence likely to influence splicing and thus might affect the translation of VDR. The frequency of this variant is lower than that of rs7975232 with 242 and 7505 homozygous mutants identified in 1000G and ExAC databases.

Importantly, genetic studies examining the role of TakI and ApaI polymorphisms in the pathogeneses of NL, DN and ESRD remained ambiguous [4‐6, 8‐12]. Considering the significance of VDR signaling in the protection against renal diseases and the ambiguity in the studies relating VDR gene polymorphism with the disease etiology, present meta-analysis comprising 2669 renal disease cases and 3342 controls was carried out to clarify the association of VDR gene TaqI and ApaI polymorphisms with nephrolithiasis, ESRD and diabetic nephropathy.

Figure 1 depicts the data extraction process for the meta-analysis. Of the 16 case-control studies retrieved on the association of TaqI polymorphism with renal disease (Table 1), four studies showed deviation from Hardy-Weinberg equilibrium [7, 13‐15]. Among the different population groups included in this meta-analysis, the largest being that of Turkish representing five case-control studies [16‐20], two studies from India [21, 22] and one each from China [23], Ireland [24], Italy [25], Spain [26] and Croatia [27]. In total, the final meta-analysis was based on the data of 2777 cases and 3522 controls representing 16 case-control studies.

Cochrane Q-test (Q: 13.72, p = 0.54) and I² (0.00) statistics showed no evidence of heterogeneity in association. Egger’s test revealed no evidence of publication bias (p = 0.14). The VDR TaqI C-allele, under allele contrast fixed effect model, was associated with renal diseases calculated collectively for DN, ESRD and NL (OR: 1.11, 95% CI: 1.03–1.20, p = 0.008). (Figure 2) As shown Table 2, subtype analysis revealed Taql C- allele to be associated with ESRD (OR: 1.17, 95% CI: 1.02–1.34, p = 0.03) (Fig. 2). Among the different ethnic groups, Turkish population showed strong association between VDR TaqI polymorphism and renal disease in allele contrast model (C vs. T, OR: 1.19, 95% CI: 1.01–1.42, p = 0.04). Sensitivity analysis revealed that omitting either of the studies had no effect on overall outcome of disease risk.

Of the 13 case-control studies (2440 cases and 3279 controls) retrieved on the association of ApaI polymorphism with renal disease (Table 3), five studies deviated from Hardy-Weinberg equilibrium [7, 15, 19, 21, 28]. Among the studies in accordance with HWE equilibrium, 3 studies were from Turkey [16, 17, 20], two from China [14, 23], and one each from Ireland [24] and Iran [29]. Cochrane Q-test (Q: 17.01, p = 0.03) and I² (48.3) statistics showed high-degree of heterogeneity in association. Egger’s test revealed no evidence of publication bias (p = 0.54). The fixed effect model showed positive association of VDR ApaI polymorphism with all the renal disease cases (C vs. A, OR: 1.10, 95% CI: 1.01–1.19), whereas, random effect model showed null association (OR: 1.05, 95% CI: 0.93–1.19) (Fig. 3). Sensitivity analysis for ApaI polymorphism revealed that the sources of heterogeneity are two studies i.e. Wang et al. and Tripathi et al. However, overall trend suggests ApaI variant as a risk factor for renal disease. As shown in Table 4, subgroup analysis revealed association of VDR ApaI polymorphism with ESRD (C vs. A, OR: 1.31, 95% CI: 1.15–1.50, p = 0.0001) and no association with NL and DN.

Deficiency of vitamin D or defective activation of VDR by its ligand, 1,25-dihydroxy vitamin D results in secondary hyperparathyroidism, angiotensin II-mediated renal damage and renal disease pathogenesis [3]. On the other hand, VDR activation suppressed inflammatory cell infiltration and inhibited nuclear factor-κB activation [30]. Likewise, active vitamin D3 and lentivirus-mediated transforming growth factor-β (TGF-β) interference effectively reduced renal fibrosis in rat models [31]. These observations highlight the importance of VDR signaling in maintaining normal renal function. Accordingly, a number of studies have investigated the effects of polymorphisms in VDR gene on renal disease etiology. Among these, TaqI, and ApaI polymorphisms are widely studied [4‐6]. However, there is a considerable ambiguity among these genetic studies, possibly stemming from sample size, ethnicity or gene-environmental interactions [4‐6, 8‐12]. To clarify whether TaqI and apaI polymorphisms have a role in renal disease pathogenesis, this meta-analysis comprising 2777 renal disease cases including DN, NL and ESRD and 3522 healthy controls was carried out. The present meta-analysis revealed an increased disease risk for subjects harboring TaqI C-allele under fixed and random effect models. Subgroup analysis based on type of renal disease showed that VDR TaqI polymorphism is associated with ESRD in allele contrast model, whereas no significant association was found between TaqI polymorphism and DN and NL. In the case of ApaI polymorphism, Apal C-allele was found to be linked to ESRD, but not with DM or NL under fixed effect model. Earlier, Yang et al. performed a meta-analysis on 1510 cases and 1812 controls and found no association of BsmI, FokI, TaqI, and ApaI polymorphisms of VDR with end-stage renal disease. Inclusion of more studies benefited the current meta-analysis.

The direct role of solar rays in the synthesis of vitamin D is well known. In human skin, solar rays facilitate the formation of vitamin D3 from 7-dehydrocholesterol, which is evident from the presence of higher mean serum vitamin D levels in summer than in winter [32]. Likewise, higher vitamin D levels were found in populations living in regions known to have longer durations of sun exposure [33].

This meta-analysis revealed the association of VDR TaqI and ApaI polymorphisms with ESRD risk. This is the first meta-analysis study to simultaneously evaluate the association of DN, NL and ESRD with renal disease risk. Ethnicity, sample size, gene-environmental interactions appear to be responsible for inconsistencies observed in the association studies examining VDR polymorphisms and renal diseases. The limitations of this meta-analysis include; exclusion of studies where raw data or full text were not accessible and one-to-one correlation between vitamin D3 profile and risk could not be established as no parallel studies were conducted.