Lilian Jara

Evidence of a Sex-Dependent Association between the MSX1 Locus and Nonsyndromic Cleft Lip with or without Cleft Palate in the Chilean Population

Prior studies have implicated an involvement of the Msx1 homeobox gene in cleft palate in mice and its homolog in humans (called MSX1 in the HOX7 gene, located on chromosome 4). In this study we present evidence of a sex-dependent association between MSX1 and nonsyndromic cleft lip/palate (NSCLP) in the Chilean population. The sample included 73 NSCLP cases, 37 from multiplex families (Mx), 36 from simplex families (Sx), and 87 controls. Polymerase chain reaction amplification of the MSX1 intragenic microsatellite (CA)n-sequence shows significant (p = 0.035) differences in the allele frequencies between NSCLP-Mx males and control males. These differences are mainly due to frequency differences in allele *2 (173 base pairs) among cases (21.9%) and controls (13.2%). When the NSCLP cases are subdivided by sex and positive family history (Mx versus Sx), the Mx males (27.8%) as well as the total NSCLP-Mx cases (25.7%) showed significantly higher frequencies of allele *2, compared to controls (11.4% and 13.2%, respectively). Analysis of the genotype data indicates that the relative risk for NSCLP is greater for persons carrying allele *2 (i.e., odds ratio [OR] larger than 1), reaching significance for all Mx cases (OR = 2.67; 95% confidence interval [CI], 1.10 to 6.52) and even more pronounced for Mx males (OR = 3.33; 95% CI, 1.08 to 10.32). Taken together, these findings support the hypothesis that the genetic variation at the MSX1 locus is a predisposing gene involved in sex-dependent susceptibility to clefting and that it also differentiates simplex from multiplex families.

Genome-wide analysis of genetic susceptibility to language impairment in an isolated Chilean population.

Specific language impairment (SLI) is an unexpected deficit in the acquisition of language skills and affects between 5 and 8% of pre-school children. Despite its prevalence and high heritability, our understanding of the aetiology of this disorder is only emerging. In this paper, we apply genome-wide techniques to investigate an isolated Chilean population who exhibit an increased frequency of SLI. Loss of heterozygosity (LOH) mapping and parametric and non-parametric linkage analyses indicate that complex genetic factors are likely to underlie susceptibility to SLI in this population. Across all analyses performed, the most consistently implicated locus was on chromosome 7q. This locus achieved highly significant linkage under all three non-parametric models (max NPL=6.73, P=4.0 × 10−11). In addition, it yielded a HLOD of 1.24 in the recessive parametric linkage analyses and contained a segment that was homozygous in two affected individuals. Further, investigation of this region identified a two-SNP haplotype that occurs at an increased frequency in language-impaired individuals (P=0.008). We hypothesise that the linkage regions identified here, in particular that on chromosome 7, may contain variants that underlie the high prevalence of SLI observed in this isolated population and may be of relevance to other populations affected by language impairments.

The sequence of eruption of the permanent dentition in a Chilean sample with Down's syndrome

The eruption of the permanent teeth in Downs individuals is reportedly delayed. The extent of such delay in comparison to normal children has been little studied. The eruption characteristics of the permanent teeth in a sample of Chilean individuals with Downs syndrome were here compared with those of the normal Chilean population. The sample consisted of 240 Downs individuals (all with trisomy 21), 116 males and 124 females. The chronological sequence of eruption in Downs children was not completely different from the normal. The least affected teeth were upper and lower first molars and central and lateral incisors. Alterations of the eruption sequence were not necessarily a consequence of alterations in the time of eruption. Asymmetries between sides of the jaw were mainly in canines and premolars. Alterations in sequence timing and asymmetry seem to be age dependent, being less frequent between 7 and 9 yr of age and more frequent between 10 and 14 yr of age. This may also reflect the larger variances of age of eruption observed in Downs individuals. Despite this, Downs children maintained a certain similarity in sequence and symmetry in comparison to normals.

Interleukin-1 gene cluster polymorphisms associated with periodontal disease in type 2 diabetes.

BACKGROUND Epidemiologic studies have shown an increased frequency, severity, and risk of periodontitis in patients with diabetes. Periodontitis is associated with certain interleukin (IL)-1 gene cluster polymorphisms. Diabetes is a proinflammatory state with increased levels of circulating cytokines suggesting a causal role for inflammation in its etiology. Common genetic factors may be involved in the susceptibility for diabetes and periodontitis. We evaluated the relationships among IL-1 gene polymorphisms, type 2 diabetes, and periodontitis. METHODS One hundred twelve patients with diabetes and chronic periodontitis, 224 patients without diabetes but with chronic periodontitis, and 208 healthy subjects without periodontitis were studied. All received a clinical periodontal examination and assessment of standard periodontal parameters. IL-1A -889, -1B +3954, and -1B -511 polymorphisms were identified by polymerase chain reaction (PCR) amplification followed by restriction enzyme digestion and gel electrophoresis. Variable numbers of IL-1RN tandem repeats were detected by PCR amplification and fragment-size analysis. RESULTS The severity and extent of periodontitis was significantly greater in patients with diabetes than in patients without diabetes. No significant differences in IL-1A -899, -1B +3954, or -1RN genotype frequencies were found between patients with diabetes and patients without diabetes. The IL-1A -889 TT genotype (odds ratio [OR] = 2.90; 95% confidence interval [CI] = 1.20 to 7.02), IL-1B +3954 TT genotype (OR = 3.54; 95% CI = 1.15 to 10.85), and IL-1B -511 CC genotype (OR = 2.10; 95% CI = 1.25 to 3.58) were significantly associated with periodontitis. The presence of an IL-1 positive genotype was significantly associated with periodontitis (OR = 1.61; 95% CI = 1.04 to 2.49). No interaction between smoking status and polymorphisms was found. CONCLUSIONS Periodontitis was significantly associated with some IL-1 gene polymorphisms. No association between diabetes and IL-1A and -1B gene polymorphisms was found.

Spectrum of BRCA1/2 point mutations and genomic rearrangements in high-risk breast/ovarian cancer Chilean families

The distribution of BRCA1/2 germline mutations in breast/ovarian cancer (BC/OC) families varies among different populations. In the Chilean population, there are only two reports of mutation analysis of BRCA1/2, and these included a low number of BC and/or OC patients. Moreover, the prevalence of BRCA1/2 genomic rearrangements in Chilean and in other South American populations is unknown. In this article, we present the mutation-detection data corresponding to a set of 326 high-risk families analyzed by conformation-sensitive gel electrophoresis and heteroduplex analysis. To determine the contribution of BRCA1/2 LGRs in Chilean BC patients, we analyzed 56 high-risk subjects with no pathogenic BRCA1/2 point mutations. Germline BRCA1/2 point mutations were found in 23 (7.1%) of the 326 Chilean families. Families which had at least three BC and/or OC cases showed the highest frequency of mutations (15.9%). We identified 14 point pathogenic mutations. Three recurrent mutations in BRCA1 (c.187_188delAG, c.2605_2606delTT, and c.3450_3453delCAAG) and three in BRCA2 (c.4969_4970insTG, c.5374_5377delTATG, and c.6503_6504delTT) contributed to 63.6 and 66.7% of all the deleterious mutations of each gene, which may reflect the presence of region-specific founder effects. Taken together BRCA1/2 recurrent point mutations account for 65.2% (15/23) of the BRCA1/2 (+) families. No large deletions or duplications involving BRCA1/2 were identified in a subgroup of 56 index cases negative for BRCA1/2 point mutations. Our study, which is the largest conducted to date in a South American population, provides a comprehensive analysis on the type and distribution of BRCA1/2 mutations and allelic variants.

SEQUENCE OF ERUPTION OF DECIDUOUS DENTITION IN A CHILEAN SAMPLE WITH DOWN'S SYNDROME

The eruption of the deciduous teeth in Downs individuals is reportedly delayed, but the extent of delay in comparison to normal children has been little studied. The eruption characteristics of the deciduous teeth in a sample of Chilean individuals with Downs syndrome were compared with those of the normal Chilean population. The sample consisted of 255 Downs individuals (all with trisomy 21), 127 males and 128 females. Boys with Downs syndrome showed significantly delayed eruption in six teeth: in the maxilla the right central incisor and right and left lateral incisors, and in the mandible the right central incisor and right and left canines. Girls with Downs syndrome showed significant delays in the eruption of 11 teeth: in the maxilla the right and left lateral incisors, right and left canines and first left molar, and in the mandible the left central incisor, right and left lateral incisors and canines and second right molar. The chronological sequence of eruption in Downs children was not completely different from that of normal individuals. With a few exceptions no significant departures from Gaussian distribution were found in the age of eruption among both normal and Downs individuals. The variance was significantly larger in cases of Downs syndrome.

Linkage Disequilibrium between MSX1 and Non-syndromic Cleft Lip/Palate in the Chilean Population

Non-syndromic cleft lip/palate (NSCLP) is a complex genetic trait. Linkage and association studies have suggested that a clefting locus could be located on chromosome 4p. Sixty Chilean families were recruited for this study; from these, we used unrelated trios to evaluate the possible linkage disequilibrium between MSX1 and NSCLP. An intragenic marker, MSX1-CA, and an extragenic marker, D4S432 at a distance of 0.8 cM from MSX1, were analyzed by means of polymerase chain-reaction with fluorescent-labeled forward primers, followed by electrophoresis on a laser-fluorescent sequencer. We carried out a transmission/disequilibrium test (TDT) for multiple alleles to evaluate the presence of linkage disequilibrium. Results showed a preferential transmission of the 169-bp allele of MSX1 (p = 0.03). Although there was no preferential transmission for the D4S432 marker, the overall extended TDT (ETDT) showed a significant result (p = 0.01). The authors’ findings support the hypothesis of the contribution of MSX1 in the etiology of NSCLP in the Chilean population.

Parent-of-origin effects for MSX1 in a Chilean population with nonsyndromic cleft lip/palate†

Based on association and sequencing studies, investigators have postulated muscle segment homeobox 1 (MSX1) as a strong candidate gene involved in the causation of nonsyndromic cleft lip with or without cleft palate (NSCLP). Parent‐of‐origin effects have been suggested for some NSCLP candidate genes but not for MSX1. The aims of the present study were to test for allele/haplotype associations applying the transmission disequilibrium test (TDT) and the transmission asymmetry test (TAT) to evaluate the possible parent‐of‐origin effects of MSX1 in Chilean patients with NSCLP. We analyzed five SNPs (rs6446693/c.‐425G>T/c.‐35G>A/rs3775261/rs12532) located from 6.3 kb upstream to 3′ UTR in a sample of 150 unrelated NSCLP case–parent trios. Four haplotypes showed overtransmission from parents to affected progeny, but individual SNPs did not. Two haplotypes presented allele combination C‐G‐A‐G (P = 0.035) and two T‐G‐C‐A (P = 0.044) (SNP order rs6446693/c.‐35G>A/rs3775261/rs12532). The rs12532 A allele had a 2.08‐fold increase in the risk of NSCLP when inherited from the father (95% CI: 1.10–4.02; P = 0.025), but not from the mother. These results could indicate epigenetic control by imprinting in the role of MSX1 in NSCLP. Different authors have proposed that some genes that play a role in NSCLP depend on parental origin. Our findings and those previously reported by our group show that a variety of factors appears to be involved in the association between MSX1 and NSCLP. The full mechanism of MSX1 in the development of NSCLP has not been fully understood.

Association between bone morphogenetic protein 4 gene polymorphisms with nonsyndromic cleft lip with or without cleft palate in a chilean population.

Nonsyndromic cleft lip with or without cleft palate (NSCLP) is one of the most common birth defects in humans with both genetic and environmental components involved in its expression. Experimental evidences have postulated that bone morphogenetic protein 4 gene (Bmp4) is involved in the etiology of cleft lip with or without cleft palate (CL/P) in mice. In our study we analyzed the association between BMP4 and NSCLP in a sample of 150 unrelated trios ascertained through affected probands. Three BMP4 polymorphisms were analyzed, two intronic (rs762642 and rs2855532) and rs1957860, located 5.7 kb upstream from BMP4. Transmission/disequilibrium tests were performed at the allele and haplotype levels. Our results did not detect preferential transmission for individual single-nucleotide polymorphisms. Significant transmission distortion was observed for haplotypes rs1957860-rs762642 (p = 0.018), especially for C-T (p = 0.015) and T-T (p = 0.018) which include the genomic region where the promoter and an enhancer of BMP4 are located. Thus, despite the positive association detected between these haplotypes and NSCLP they probably do not have a functional effect on BMP4 expression or protein activity but possibly reflect NSCLP susceptibility changes which are in linkage disequilibrium with these polymorphisms. The findings of our study support a role for BMP4 in NSCLP in the admixed Chilean population.

Association Between 10 Microsatellite Markers and Nonsyndromic Cleft Lip Palate in the Chilean Population

Objective The objective of this case-control study was to evaluate the possible association between nonsyndromic cleft lip/palate (NSCLP) and 10 genetic markers in four chromosomal regions in the admixed Spanish-Amerindian Chilean population. Setting Study participants included 56 patients with NSCLP identified and interviewed for positive family history during the course of clinical examinations at different rehabilitation centers in the cities of Santiago and Talca, Chile. A control group of 59 normal individuals without known familial antecedents of clefting was obtained from blood bank donors of the University Hospital, University of Chile. Cases and controls belonged to low- to low-middle socioeconomic strata. Results Ten markers from chromosome 4p, 4q, 6p, 17q, and 19q were assessed (MSX1, D4S175, D4S192, F13A1, EDN1, D6S89, D6S105, D6S109, D17S579, BCL3). Four of them showed significant deviations from Hardy-Weinberg expectations in controls, according to the exact test (D4S192, BCL3, F13A1, and D6S89). The case-control comparison by means of the CLUMP program showed significant differences only in BCL3, and D6S109 almost reached statistical significance. Conclusions Most of the genetic regions with positive results in Caucasian populations may not be involved in NSCLP in Chile, regardless of the positive evidence for the candidate region on chromosome 19. Similar findings have been reported recently in the Chinese population.