Comparative evolutionary pharmacogenetics of CYP2D6 in Ngawbe and Embera Amerindians of Panama and Colombia: role of selection versus drift in world populations.

The development of CYP2D6 has been attributed to the need of earth-dwelling animals to detoxify toxic xenobiotics (phytoalexins) present in plants. This hypothesis has been extrapolated to humans, but is yet unconfirmed. Therefore, we studied two Amerindian populations as the best available model to test the effect of selection through diet on human CYP2D6 evolution. The frequency of sparteine poor metabolizers in Ngawbe was 4.4% (n = 344), while the frequency in Embera was 2.2% (n = 153). Among Ngawbe and Embera, CYP2D6*4 (allelic frequencies for each tribe, respectively: 0.171; 0.14), CYP2D6*6 (0.005; 0.011) and CYP2D6*10 (0.175; 0.069) were detected, while CYP2D6*3, CYP2D6*5, CYP2D6*9 and CYP2D6*16 were absent. All poor metabolizers possessed either CYP2D6*4 or CYP2D6*6 and there were no disagreements between genotypic and phenotypic data. The total frequency of mutant alleles showed no difference among Amerindians or when compared to Caucasians. It was higher than in Chinese, since the frequency of CYP2D6*4 was higher in Amerindians. XbaI restriction fragment length polymorphisms haplotypes were very homogeneous in Amerindians, because the only fragment that hybridized with the CYP2D6 cDNA probe was the 29 kb (not 42/44 kb or 11.5/13 kb). This indicated no gene cluster recombinations that generate insertions or deletions. We propose that in earlier hominids and humans, CYP2D6 had increasingly become a vestigial characteristic unconstrained by dietary stressors, as a result of cultural survival strategies. Human CYP2D6 evolution was preferentially affected by random genetic drift, and not by adaptive or purifying selection.

Isoniazid Acetylation Phenotyping in the Japanese: The Molar Metabolic Ratio INH/AcINH.

The N-acetylation phenotyping of isoniazid (INH) was studied in 434 unrelated Japanese pulmonary tuberculosis patients. The frequency of slow acetylators was determined using three methods based on the urinary levels of INH and AcINH: percentage acetylisoniazid (%AcINH), the inactivation index or acetylation index (AcINH/INH), and the molar metabolic ratio (INH/AcINH) in urine. Frequency histograms and probit plots were constructed with the data obtained from each method. Using %AcINH with the conventional antimode of 70%, the number of slow acetylator was 12.7%. Using AcINH/INH, the number of slow acetylator was 52% according to the conventional antimode of 6.0. The molar metabolic ratio INH/AcINH showed explicitly the best bimodality and a clear-cut antimode among these three methods. From probit plots of INH/AcINH, an antimode of 0.45 can be suggested for the 434 Japanese patients; 377 patients (86.9%) as rapid acetylator and 57 patients (13.1%) as slow acetylator.

[Human biodiversity and its effects on the pharmacological variability: CYP2D6 and NAT2 enzymes in Amerind populations of Panama, Colombia and Costa Rica]. / La biodiversidad humana y sus efectos sobre la variabilidad farmacologica: enzimas CYP2D6 y NAT2 en poblaciones amerindias de Panama, Colombia y Costa Rica.

Human biodiversity originates partially from human microevolution, which have produced different populations. This biodiversity is responsible for most of the variability in drug response. We present the methodology employed in population pharmacology studies and general information about the CYP2D6 and NAT2 systems. We report results obtained in Embera and Ngawbe Amerindians, who are characterized by a low phenotypic and genotypic CYP2D6 diversity. In regard to NAT2, Amerindians are distinguished by a high allelic frequency of S3 and low ones of S1 and S2, situation which is reversed in Caucasians.

La biodiversidad humana y sus efectos sobre la variabilidad farmacologica: enzimas CYP2D6 y NAT2 en poblaciones amerindias de Panama, Colombia y Costa Rica / Human biodiversity and its effects on the pharmacological variability: CYP2D6 and NAT2 enzymes in Amerind populations of Panama, Colombia and Costa Rica

Human biodiversity originates partially from human microevolution, which have produced different populations. This biodiversity is responsible for most of the variability in drug response. We present the methodology employed in population pharmacology studies and general information about the CYP2D6 and NAT2 systems. We report results obtained in Embera and Ngawbe Amerindians, who are characterized by a low phenotypic and genotypic CYP2D6 diversity. In regard to NAT2, Amerindians are distinguished by a high allelic frequency of S3 and low ones of S1 and S2, situation which is reversed in Caucasians

Reduced mtDNA diversity in the Ngöbé Amerinds of Panamá.

Mitochondrial DNA (mtDNA) haplotype diversity was determined for 46 Ngöbé Amerinds sampled widely across their geographic range in western Panamá. The Ngöbé data were compared with mtDNA control region I sequences from two additional Amerind groups located at the northern and southern extremes of Amerind distribution, the Nuu-Chah-Nulth of the Pacific Northwest and the Chilean Mapuche and from one Na-Dene group, the Haida of the Pacific Northwest. The Ngöbé exhibit the lowest mtDNA control region sequence diversity yet reported for an Amerind group. Moreover, they carry only two of the four Amerind founding lineages first described by Wallace and coworkers. We posit that the Ngöbé passed through a population bottleneck caused by ethnogenesis from a small founding population and/or European conquest and colonization. Dating of the Ngöbé population expansion using the Harpending et al. approach to the analysis of pairwise genetic differences indicates a Ngöbé expansion at roughly 6800 years before present (range: 1850-14,000 years before present), a date more consistent with a bottleneck at Chibcha ethnogenesis than a conquest-based event.

Evolutionary pharmacogenetics of CYP2D6 in Ngawbe Guaymi of Panama: allele-specific PCR detection of the CYP2D6B allele and RFLP analysis.

Gene cluster CYP2D controls the biosynthesis of enzyme CYP2D6, which is responsible for the polymorphic oxidation of sparteine, debrisoquine and related drugs. This cluster consists of the functional gene D6 and of two pseudogenes, D7 and D8. RFLP Bam HI analysis of CYP2D in 37 unrelated and eight related Ngawbe Guaymi Amerindians of Panama showed a polymorphism characterized by the presence of two alleles: 4.7 + 7.9 and 2.3 + 6.0 (frequencies: 0.63 and 0.37, respectively, n = 35 unrelated subjects). The possible genotypes for these alleles follow the Hardy-Weinberg distribution (chi 2 = 1.76; 0.10 < p < 0.25). All PMs of sparteine or debrisoquine (n = 7) were homozygotes for the second allele, but not all homozygotes (n = 10) were PMs, so there was not an exclusive association between the Bam HI genotype and the observed phenotype. A similar analysis with the endonuclease Xba I proved to be non-informative in relation to phenotype, since all subjects (n = 40) showed only the 29 kb allele. Allele-specific PCR studies of selected subjects indicated the existence of the CYP2D6B allele (freq = 0.17; C.I.95% = 0.085, 0.29; n = 30 unrelated subjects), in addition to the wild-type. The mutant CYP2D6B allele was responsible for the enzyme deficiency present in PMs. Its presence in Amerindians suggests that this allele has a far more ancient evolutionary history than previously thought. The over-all RFLP and PCR analyses point to a diminished genetic diversity for the Ngawbe subjects, consistent with their demographic history and population genetics.

["Cholos de Coclé": determination of their racial mixture and genetic origins]. / Estudio sobre los "cholos de Coclé": determinación de su mezcla racial y orígenes genéticos.

We have found, in this first genetic study of a supposedly admixed Panamanian population, that the cultural group known as "cholos of Coclé" constitute a trihybrid mixture, whose genetic pool has the following composition: 44% Amerindian, 38% Caucasoid and 18% Negroid. Similarly, we have detected Amerindian genes, such as LDHB--Gua and TFchi, in proportions that relate this population with the extant Ngawbé (Guaymí). Nevertheless, the very high frequency of variant PEPA--KUN seems to indicate the genetic contribution of Amerindian populations from Eastern Panama, possibly from the extinct indigenous group cueva. This variant is frequently found among the present-day Kuna, but has not been detected among Nagawbé and Buglé.

Debrisoquine polymorphism: novel CYP2D6 gene Bam HI restriction fragment length polymorphism in the Ngawbé Guaymí Indian of Panama.

Markedly decreased cytochrome P450-mediated metabolism of debrisoquine, sparteine, and more than two dozen additional commonly prescribed drugs is an autosomal recessive trait that has been associated with several RFLP patterns involving the CYP2D6 gene. In Caucasians there are at least six variant alleles known to be correlated with the 'poor metabolizer' (PM) phenotype. We examined debrisoquine and sparteine metabolism and CYP2D6 RFLP patterns in 22 Ngawbé Guaymí Indians of Panama. We studied a two-generation family, a three-generation family, and three other unrelated PM individuals. Digestion of all 22 DNA samples with Xba I or Hind III did not produce the same varying CYP2D6 RFLP patterns as those commonly seen in at least two-thirds of all Northern European Caucasians and Chinese so far screened. In contrast, we found a single heretofore undescribed Bam HI polymorphism that was correlated with the PM phenotype among all Ngawbé Guaymí individuals examined. It is possible that this novel RFLP might represent a recent founder effect that has occurred in this unadmixed Amerindian tribe within the past 20,000-30,000 years.

Unimodal distribution of the metabolic ratio for debrisoquine in Cuna Amerindians of Panama.

1. The metabolic oxidation of debrisoquine (DB) was studied in 89 non-related Cuna Amerindian subjects. 2. Means and standard deviations for urinary recoveries of the intact drug and its 4-hydroxy metabolite (4-HD) were: %DB: 6.8 +/- 4.5; %4-HD: 16.0 +/- 9.1; %sum: 22.8 +/- 12.0. The log10 metabolic ratios for DB (LMRDB) were distributed within a single mode of insignificant skewness (-0.01, P greater than 0.10), which was unimodal (log kernel density and maximum likelihood methods) and normal (chi 2 = 22.5; d.f. = 15; P greater than 0.09; power of the test greater than 80%). 3. Therefore, no poor metabolizers (95% C.I.: 0.1%, 5.2%) were detected in the population sample studied.

[Population pharmacology and ethnopharmacokinetics in 3 Amerindian groups from Panama: Cuna, Ngawbe Guaymi and Teribe]. / Farmacología poblacional y etnofarmacocinética de tres grupos amerindios de Panamá: Cuna, Ngawbe guaymi y Teribe.

We report on studies of the oxidative routes associated with the polymorphic metabolism of debrisoquine, sparteine and mephenytoin, as well as on the pathway catalyzed by N-acetyl-transferase. Normal, healthy non-related subjects were studied: 250 Cuna, 285 Ngawbé Guaymí and 20 Teribe. These studies were aimed at establishing differences or similarities among Amerindian tribes and between them and Caucasians, since certain abnormal or non-expected results in clinical response to drugs could be due to racial differences. In those cases, it was considered necessary to evaluate the degree of toxicity and danger of drugs administration, and their effective therapeutic actions.

Microevolution in lower Central America: genetic characterization of the Chibcha-speaking groups of Costa Rica and Panama, and a consensus taxonomy based on genetic and linguistic affinity.

There is evidence that Amerindians have continuously occupied the lower Central American Isthmus for as long as 10,000 years. There remains some doubt about the relationships of these original colonizers to the resident peoples of this zone at the time of European contact (approximately A.D. 1500). We present new genetic data for up to 48 genetic loci for 570 members of six Chibcha-speaking tribes of lower Central America--the Boruca, Bribri, Cabecar, and Guatuso of Costa Rica and the Kuna and Teribe of Panama--and delineate the genetic affinities among the various groups (these six tribes and the Guaymi and Bokota) of lower Central America. We convert standard genetic distance metrics into a form that is linear with the effective time since divergence, and we compare the genetic distances with linguistic distances for the same groups (r = .74, P less than .001). Geographic affinity accounts for some of the genetic divergence among groups (r = .49, P less than .084) and for some of the linguistic divergence (r = .53, P less than .037), but the correspondence between geographic position and taxonomic affinity is not high. We combine all of the genetic and linguistic data to construct a synthetic overview taxonomy of the lower Central American Chibcha. Both the genetic and linguistic data exhibit hierarchical organization of tribal groups, showing a general east-to-west pattern of grouping, with greater affinities between close neighbors. The presence of private genetic variants of some antiquity within the region and their absence outside the zone, coupled with the essential absence of the DI*A polymorphism of mongoloid origin that is widespread outside the zone, argue for a relatively isolated development of the Central American Chibcha. Our results do not support the old view of lower Central America as a frontier between more advanced cultures to the north and south. Any such explanation would require recent waves of migration from outside the region, migration that is not compatible with either the genetic or linguistic data or with the archaeological history of the region.

A preliminary note on the transient polymorphic oxidation of sparteine in the Ngawbé Guaymí Amerindians: a case of genetic divergence with tentative phylogenetic time frame for the pathway.

The oxidation of sparteine was studied in a total of 121 Ngawbé Guaymí volunteers in Panama, 97 of whom were unrelated. When presented in a frequency histogram, the results of the log10 of the metabolic ratios (LMR) indicated the existence of two modes, the largest of which exhibited a normal distribution (alpha = 0.05; chi 2 = 5.46). A preliminary assignment of an antimode for this population sample is proposed, located within the region of LMR 0.65 to 0.85 vs LMR of 1.3 for white subjects, and results in five poor metabolizers (PMs) (5.2%). This is in contrast to the absence of PMs (0/210) we have reported for the Cuna Amerindians. The microevolution of the sparteine route, corresponding to a tenfold change in the frequency of PMs, is likely to have occurred within their genetic divergence time. These observations of the divergence of a metabolic route of therapeutic importance and the proposal of a time frame for its microevolution constitute the first cases in the literature.

The oxidative metabolism of sparteine in the Cuna Amerindians of Panama: absence of evidence for deficient metabolizers.

Sparteine sulfate (50 mg) was administered to 170 Cuna Amerindians, 142 of whom were unrelated, and the drug and its dehydrometabolites were determined in the 0- to 12-hour urine samples. The log10 of the metabolic ratio was unimodally, but not normally, distributed and showed the following values: mean -0.21 +/- 0.26, median -0.24, limits -0.73 and 0.76, skewness 1.00, and kurtosis 4.95. On the basis of these results, it can be concluded that there are no deficient metabolizers in the Cuna sample population studied. However, the similarity of the skewness found between the Cuna sample population studied and the extensive Canadian white group, as well as an inflection point at 6.3 U in the former's probit plot, suggests the existence of at least two subgroups congregating within the same single mode in the frequency distribution curve. The use of the inflection point is discussed thoroughly, concluding that although it does not allow exclusion of the existence of genotypically different subgroups, the limitations of the data do not permit its use to determine the number of heterozygotes and thus the existence of polymorphism. The possibility of an isozyme variant, consistent with the general genetic structure of Amerindians, as suggested by the coexistence of two subgroups within the unimodal curve, is entertained.

Mephenytoin hydroxylation in the Cuna Amerindians of Panama.

1 Mephenytoin p(4')-hydroxylation, which is deficient in 3-5% of Caucasians, was examined in 96 Cuna Amerindians of Panama. 2 Attempts were made to exclude poor compliance with urine collection and ingestion of the drug dose since the assignment of phenotype was based upon urinary recovery of the metabolite. These involved the measurement of the urinary recovery of sparteine, added to the ingested capsule, and of the renal excretion of creatinine. 3 Of the 90 Cunas deemed to be reasonably complaint, none of them appeared to be deficient in p(4')-hydroxylation of mephenytoin.

On phenotyping with isoniazid: the use of urinary acetylation ratio and the uniqueness of antimodes. Study of two Amerindian populations.

Various conventions have been used to express the activity of polymorphic hepatic N-acetyltransferase of isoniazid. Among them, two of the most common are the "percentage of acetylisoniazid" and the "inactivation index." A third alternative convention is proposed, the "molar acetylation ratio," which showed the most clear-cut bimodal distribution when applied to the results obtained for Cuna and Teribe Amerindians living in Panama. Through this method a unique antimode was assigned to each Amerindian group and the same frequency of slow acetylators (24% to 29%) was found, unlike the results obtained by conventional approaches and antimodes derived from white populations.