Sickle cell trait and targeted genomic variants in chronic kidney disease an African cohort
Background Chronic Kidney Disease (CKD), has a high and increasing burden in sub-Saharan Africa. Environmental factors that have been associated to CKD are associated with multiple co-morbidities such as hypertension, diabetes, and HIV. Some genetics factors such APOL1 have been associated with the highest burden of CKD among population of African ancestries. Other emerging genetic factors such as Sickle Cell trait (SCT) have been investigated mostly among African Americans. Sickle Cell trait (SCT) has the highest burden in sub-Saharan Africans, because of a natural selection, attributed to its protective advantages against the severest form of Malaria, caused by Plasmodium falciparum. Many studies showed that SCT has an impact on the normal functioning of the kidneys among African Americans with some studies indicating significant association between SCT and CKD. However, no study has been reported from Sub-Saharan Africa, where most SCT carrier reside. Moreover, there are multiple other loci and variants in the genome that have been associated with CKD in many populations, and that are used for Polygenic Risk Score (PRS) models but have not been explored in populations living in Africa. Aims This project aimed to study in a sub-Saharan African cohort, the association between 1) Sickle cell trait (SCT) with Chronic Kidney disease (CKD), and 2) the association of CKD with 29 targeted single nucleotide polymorphisms (SNPs) identified in multiple Genome-Wide Association studies (GWAS). Methods Patients and controls: 300 Cameroonian adult participants were included: 150 CKD cases and 150 non-CKD age, sex, and comorbidities matched controls. Molecular methods: SCT heterozygosity was determined by RFLP-PCR using the restriction enzyme DdeI. A total of 29 targeted SNPs was genotyped using MassArray and TaqMan techniques, followed by Sanger sequencing in a subset of samples. 11 Statistical Analysis: Descriptive statistics and logistic regression, and Fisher exact test were used. Functional pathway analysis: following the identification SNPs with significant association with CKD, we performed functional pathway test using the Linux programme Cytoscape. Results The mean age of cases was 53 years (range 46-55 years), with 43% that were female; there were no age and sex significant differences with controls. We identified, an expected, association between CKD and various co-morbidities, demographic and anthropometric variables: hypertension (p value = 5.16X10-9 ), HIV (p value = 2.68x10- 9 ), diabetes (p value = 7.12X10-7 ), BMI (p value = 4.58X10-8 ) and age (p value = 4.5X10-8 ). HbAS carrier status was significantly associated CKD (p value= 4.3X10-9 ; Odds Ratio:7.05). Only three targeted SNPs (3/29) previously associated with CKD in GWAS among African Americans, European and Asian population, were significantly associated with CKD among this group of Cameroonians (KBTBD2 rs3750082, PTPRO rs7956634 and LPR2 rs4667594 with p values of 0.02335, 0.0408 and 0.0398). Genes protein-protein interactions analysis identified the two key functional pathways and one network cluster that could play a crucial role in kidney dysfunctions. Lastly, we distinguished that HbS carrier state doesn’t influence the relationship between APOL1 G1/G2 risk alleles and CKD (p value = 0.5725) in this group from subSaharan Africans. Conclusion and perspectives Our study illustrates a strong association between SCT and CKD, an important discovery that will have a major implication in preventative medicine policies and practices in both sub-Saharan African where there is a very high prevalence of SCT. The data also has global resonance, with the projected increase in the prevalence of 12 individual with SCT, due to migration and the improve life expectancy and genetic fitness of people living with both SCT and SCD. We identified a relatively low proportion of (3/29) of target SNPs positively associated with CKD among this group of Cameroonians. The study illustrates that the vast majority of targeted SNPs associated with CKD in GWAS studies in multiple populations including African American, Europeans, and Asians, are not relevant for sub-Saharan Africans, indicating the urgent need to include diverse populations, specifically those living in Africa. Therefore, the data support the possible bias in currently available Polygenic Risk Score generated from GWAS data, where population from sub-Saharan Africa are largely underrepresented. The data further indicate that there is potential to discover new loci associated with CKD when investigating populations of African ancestry living in Africa.
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