Supplementary information: Ethnic variations in the Brazilian

Supplementary information: Ethnic variations in the Brazilian population
It is widely recognized that population stratification may be a confounding factor in
genetic association studies and the impact of stratification in population-based association
studies has been discussed extensively (1-3). With respect to the polymorphisms analyzed
here, especially PIN3 and PEX4, previous studies demonstrate that allelic and genotypic
frequencies are distributed differentially according to ethnic background. For instance, in a
case-control study performed by Weston et al. (1997) on the association of TP53
polymorphisms and breast cancer risk in individuals from the ethnically heterogeneous
population of New York city, PEX4 and PIN3 allele frequencies were significantly different
between Caucasian, Hispanic and African-american individuals (4). These differences were also
shown by Marcel et al. (2010) who analyzed 90 Caucasian, Asian and African individuals from
the Hapmap dataset (5). Thus, it is be reasonable to consider that population structure
interferes with association studies that involve these polymorphisms, in particular in the case
of populations reputed as ethnically diverse and heterogeneous such as the population of
In the context of the present study, however, both the information available on the
population structure in Southern Brazil and our data on phenotypic and genotypic variations
among the subjects we have recruited, support that the impact of population structure on
allele distributions and associations are, at the most, limited, and do not significantly bias our
results. This interpretation is based on the following considerations:
1. Landmark studies conducted with a large number of Brazilian subjects have concluded that
the current Brazilians have contributions of different continental ancestries (European,
Amerindian and African). However the proportion of the contribution of these parental stocks
varies significantly among different Brazilian regions. In one of such recent molecular analyses,
Pena and collaborators (2011) demonstrated that European ancestry was predominant in all
Brazilian regions, with proportions ranging from around 60% in the Northeast to around 80% in
the South (6), corroborating previous studies that show that populations from Southern Brazil
have a major European contribution (7-10). Thus, Southern Brazil (including the State of Rio
Grande do Sul and the city of Porto Alegre, where the study was conducted), has the largest
proportion of European contribution in the country. This has been demonstrated in several
studies, where populations from this region have been described as having “transplanted
European genomes” (9). Furthermore, even in individuals who self-declare themselves as
“Black”, the genomic European contribution is highly significant (6, 7). This first point argues in
favor of a less heterogeneous study population with a predominance of European genomes
and ancestry, as defined by population-based studies
2. When analyzing self-reported skin color, which in Brazil is commonly used as surrogate to
define an equivalent of “race” or ethnic background (6, 11), patients were separated in two
groups: “white” (women who defined themselves with Caucasian features) and “non-white”
(women that identified themselves with features suggestive of some level of African ancestry,
such as “mulato” or “pardo”). No significant difference in the distribution of self-denominated
skin color was observed among the four study subgroups. As expected from previous
population-based studies in the region of Porto Alegre, Rio Grande do Sul, the majority of
individuals in all four groups self-denominated them as “white”, corroborating with the
previous assumptions described in argument 1 (see Table S1).
3. When analyzing allele frequencies of TP53 polymorphisms PIN3 and PEX4, no significant
differences were observed between any of the 4 subgroups of the present study (IVF, END,
Fertile and Unselected) and allele frequencies reported in other studies on European
populations (see Table S3).
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