Brazilian multicenter study of association between polymorphisms in CRISPLD2 and JARID2 nonsyndromic oral cleft

Orientador: Ricardo Della ColettaTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: Variantes nos genes CRISPLD2 (cysteine-rich secretory protein LCCL domain containing 2) e JARID2 (jumonji, AT-rich interaction domain 2) foram associadas com um risco aumentado para o desenvolvimento de fissuras do lábio e/ou palato não-sindrômica (FL/PNS). Neste estudo caso-controle nós analisamos a possível associação de polimorfismos de nucleotídeo único (SNPs) nos genes CRISPDL2 e JARID2 na população brasileira com FL/PNS. Como a predisposição genética para as FL/PNS é dependente da composição étnica, nós realizamos uma análise estruturada levando em conta a variação da ancestralidade genômica de cada indivíduo. Quatro polimorfismos no gene CRISPDL2 (rs1546124, rs8061351, rs2326398 e rs4783099) e 4 no gene JARID2 (rs6915344, rs2299043, rs2237138 e rs2076056), previamente associados com FL/PNS, foram genotipados em 785 pacientes brasileiros com FL/PNS (549 com fissura labial com ou sem fissura palatina-FL±PNS e 236 com fissura palatina isolada-FPNS) e 693 indivíduos clinicamente normais. A distribuição genotípica de todos os SNPs no grupo controle respeitou o equilíbrio de Hardy-Weinberg, exceto o SNP rs6915344 no gene JARID2 que foi excluído das análises posteriores. Após o ajuste para as diferenças na ancestralidade genômica, associações alélicas entre o polimorfismo rs2237138 no gene JARID2 e uma diminuição no risco para FL±PNS (OR: 0,80; 95% IC: 0,67-0,97; p=0,02) e entre o alelo T do polimorfismo rs4783099 no gene CRISPDL2 e um aumento no risco para FPNS (OR: 1,31; 95% IC: 1,05-1,62; p=0,01) foram observadas. Os SNPs no gene CRISPDL2 não demonstraram desequilíbrio de ligação, enquanto que os SNPs no gene JARID2 revelaram um forte desequilíbrio de ligação entre eles. Nossos resultados sugerem que o SNP rs2237138 no gene JARID2 apresenta um efeito protetor contra FL±PNS, enquanto que o SNP rs4783099 no gene CRISPLD2 representa um fator de risco para FPNS na população brasileiraAbstract: Variants in the cysteine-rich secretory protein LCCL domain containing 2 gene (CRISPLD2) and in the jumonji, AT-rich interaction domain 2 gene (JARID2) were previously shown to influence nonsyndromic oral cleft susceptibility. Herein, we performed a case-control study to examine the potential association of single nucleotide polymorphisms (SNPs) in CRISPLD2 and JARID2 with nonsyndromic cleft lip and/or palate (NSCL/P) in the Brazilian population. Given the ethnicity-dependent genetic predisposition to NSCL/P, we performed a structured analysis taking into account the genomic ancestry variation of each individual. Four SNPs in CRISPLD2 (rs1546124, rs8061351, rs2326398 and rs4783099) and 4 in JARID2 (rs6915344, rs2299043, rs2237138 and rs2076056), that were previously reported to be associated with NSCL/P, were genotyped in 785 Brazilian patients with NSCL/P (549 with cleft lip with or without cleft palate-NSCL±P, and 236 with cleft palate only-NSCPO) and 693 unaffected Brazilian controls. Genomic ancestry was assessed with a set of 40 biallelic short insertion/deletion variants previously validated as ancestry informative markers of the Brazilian population. After adjustment of ancestry variations, allelic analysis revealed marginal associations between the CRISPLD2 rs4783099 T allele and increased risk for NSCPO (OR: 1.31, 95% CI: 1.05-1.62, p=0.01) and between JARID2 rs2237138 and decreased NSCL±P risk (OR: 0.80, 95% CI: 0.67-0.97, p=0.02). No linkage disequilibrium was found between CRISPLD2 SNPs, while JARID2 SNPs were in complete linkage disequilibrium. Our results suggest that CRISPLD2 rs4783099 may represent a risk factor for NSCPO while JARID2 rs2237138 shows a protective effect against NSCL±P in the Brazilian populationDoutoradoPatologiaDoutora em Estomatopatologia33003033009P4CAPESCNP

Extracellular vesicles derived from cancer-associated fibroblasts induce the migration and invasion of oral squamous cell carcinoma

As one of the most abundant constituents of the tumour microenvironment (TME), cancer-associated fibroblasts (CAF) display critical roles during tumour progression and metastasis. Multiple classes of molecules including growth factors, cytokines, proteases and extracellular matrix proteins, are produced by CAF to act as mediators of the stroma-tumour interactions. One of the main channels for this communication is associated with extracellular vesicles (EV), which are secreted particles loaded with protein and genetic information. In this study, we evaluated the effects of EV derived from CAF primary human cell lines (n = 5) on proliferation, survival, migration, and invasion of oral squamous cell carcinoma (OSCC) cells. As controls, EV from human primary-established normal oral fibroblasts (NOF, n = 5) were used. Our in vitro assays showed that CAF-EV significantly induces migration and invasion of OSCC cells and promote a disseminated pattern of HSC-3 cell invasion in the 3D organotypic assay. Furthermore, gene expression analysis of EV-treated cancer cells revealed changes in the pathways associated with tumour metabolism and up-regulation of tumour invasion genes. Our findings suggest a significant role of CAF-EV in promoting the migration and invasion of OSCC cells, which are related to the activation of cancer-related pathways.Peer reviewe

Analysis of the correlation between periodontal status and polymorphisms in the IL-1, IL-6, TNF-alpha and paraoxonase enzyme in individuals affected or not with Diabetes type I and II

Orientador: Rebeca de Souza AzevedoDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: A doença periodontal (DP) é uma doença inflamatória crônica de ampla distribuição etária e mundial, que resulta da interação entre fatores etiológicos heterogêneos e que pode sofrer influência ambiental e sistêmica, especialmente na presença do diabetes melito (DM). Os mecanismos que envolvem a associação entre o DM e a presença da DP ainda não estão completamente elucidados, mas as características genéticas ou epigenéticas do hospedeiro, como os polimorfismos genéticos, podem influenciar esta relação. Dessa forma, o objetivo deste estudo foi avaliar o envolvimento de polimorfismos nos genes de IL1-? (-889), de IL1-? (+3984), de IL-6 (-174), de TNF-? (-308) e da paraoxanase (-192) e da paraoxonase (-55) no desenvolvimento de DP e DM em um grupo de pacientes brasileiros. Os polimorfismos rs1800587 e rs1143634 no gene IL-1, rs1800796 no gene IL6, rs1800629 no gene TNF-?, e rs662 e rs854560 no gene paraoxanase foram genotipados em 302 adultos, dos quais 96 eram diabéticos com DP (ADM+DP), 20 eram diabéticos com saúde periodontal (ADM-DP), 112 eram normoglicêmicos com DP (AN+DP) e 74 eram normoglicêmicos com saúde periodontal (AN-DP); e em 88 crianças e adolescentes, dos quais 11 eram diabéticos com gengivite (CDM+G), 15 eram diabéticos com saúde gengival (CDM-G), 30 eram normoglicêmicos com gengivite (CN+G) e 32 eram normoglicêmicos com saúde gengival (CN-G). No grupo de pacientes adultos, a presença do alelo T no polimorfismo rs1143634 no gene IL1-? foi mais frequente nos indivíduos ADM+DP que nos indivíduos AN-DP, gerando um OR de 14,9 (95% IC: 8,6-25,9; p<0,0001); a presença do alelo A no polimorfismo rs1800629 no gene TNF-? foi mais frequente nos indivíduos ADM+DP que nos indivíduos AN-DP, gerando um OR de 22,7 (95% IC: 6,96 -74,5 p<0,0001), e a presença do alelo C no polimorfismo rs1800796 no gene IL6 foi mais frequente nos indivíduos ADM+DP que nos indivíduos AN+DP, gerando um risco de recorrência de 2,38 (95% IC:1,61-3,5; p<0,0001), e que nos indivíduos AN-DP, gerando risco de recorrência de 2,87 (1,79-4,5; p<0,0001). No grupo de pacientes crianças e adolescentes, a presença do alelo C no polimorfismo rs1800796 no gene IL-6 foi mais frequente em indivíduos CN+G que nos indivíduos CN-G, gerando um risco de recorrência de 121,1 (95% IC:32,3-145,3; p<0,0001), e foi mais frequente nos indivíduos CDM+G que nos indivíduos CN-G, gerando um risco de recorrência de 104 (95% IC:11,4-136,6; p<0,0001). Os resultados deste estudo evidenciam a associação dos polimorfismos rs1143634 no gene IL1-?, rs1800796 no gene IL6 e rs1800629 no gene TNF-? com a suscetibilidade genética ao desenvolvimento de DP e/ou DM na população adulta estudada; e a associação do polimorfismo rs1800796 no gene IL-6 com a suscetibilidade genética ao desenvolvimento de DP e/ou DM na população de crianças e adolescentes estudadaAbstract: Periodontal disease (PD) is a chronic inflammatory disease of worldwide distribution that may affect all age groups, and results from the interaction between heterogeneous etiologic factors, such as environmental and systemic influence, especially in the presence of diabetes mellitus (DM). Mechanisms involving the association between DM and the presence of PD are still not fully elucidated, but the genetic or epigenetic alterations of the host, such as the genetic polymorphisms may influence this relationship. Thus, the aim of this study was to evaluate the involvement of polymorphisms in the genes of IL1-? (-889), IL1- (+3984), IL-6 (-174), TNF-? (-308) and paraoxanase (-192) and paraoxonase (-55) in PD and/or DM development in a group of Brazilian patients. The polymorphisms rs1800587 and rs1143634 from IL1 gene, rs1800796 form IL6 gene, rs1800629 from TNF-? gene, and rs662 and rs854560 from paraoxanase gene were genotyped in 302 adults, of whom 96 were diabetic with PD (ADM+PD), 20 were diabetics with periodontal health (ADM-PD), 112 were normoglycemic with PD (AN+PD) and 74 were normoglycemic with periodontal health (AN-PD); and in 88 children and adolescents, of whom 11 were diabetic patients with gingivitis (CDM+G), 15 were diabetics with gingival health (CDM-G), 30 were normoglycemic with gingivitis (CN+G) and 32 were normoglycemic with gingival health (CN-G). In the group of adult patients, the presence of T allele at the rs1143634 polymorphism in the IL1-? gene was more frequent in ADM+PD than in AN-PD, generating an OR of 14, 9 (95% CI :14,9-25, 9; p <0.0001), the presence of A allele at the rs1800629 polymorphism in the TNF-? gene was more frequent in ADM+PD than in AN+PD, generating an OR of 22,7 (95% CI 6.96 -74.5; p <0.0001), and the presence of C allele at the rs1800796 polymorphism in the gene IL6 was more frequent in ADM+PD than in AN+PD, generating an OR of 2.38 (95% CI :1,61-3, 5; p <0.0001), and it was more frequent in ADM+PD than in AN-PD, generating an OR of 2.87 (1.79 to 4.5, p <0.0001). In the group of children and adolescents, the presence of C allele at the rs1800796 polymorphism in the IL6 gene is more frequent in NC+G than in NC-G, generating an OR of 121.1 (95% CI :32,3-145, 3; p <0.0001), and it was more frequent in CMD+G than in NC-G, generating an OR of 104 (95% CI :11,4-136,6; p <0.0001). The results of this study show the association of rs1143634 polymorphisms in the IL1-? gene, rs1800796 in the IL6 gene, and rs1800629 in the TNF-? gene with genetic susceptibility to PD and/or DM development in the adult population studied, and association of the polymorphism rs1800796 in the IL6 gene with genetic susceptibility for PD and/or DM development in the children and adolescents population studiedMestradoEstomatologiaMestra em Estomatopatologi

Brazilian multicenter study of association between polymorphisms in CRISPLD2 and JARID2 and non-syndromic oral clefts

Variants in the cysteine-rich secretory protein LCCL domain containing 2 gene (CRISPLD2) and in the jumonji, AT-rich interaction domain 2 gene (JARID2) were previously shown to influence non-syndromic oral cleft susceptibility. Herein, we performed a case−control study to examine the potential association of single-nucleotide polymorphisms (SNPs) in CRISPLD2 and JARID2 with non-syndromic cleft lip and/or palate (NSCL/P) in the Brazilian population. Given the ethnicity-dependent genetic predisposition to NSCL/P, we performed a structured analysis taking into account the genomic ancestry variation of each individual. Four SNPs in CRISPLD2 (rs1546124, rs8061351, rs2326398, and rs4783099) and four in JARID2 (rs915344, rs2299043, rs2237138, and rs2076056), that were previously reported to be associated with NSCL/P, were genotyped in 785 Brazilian patients with NSCL/P (549 with cleft lip with or without cleft palate-NSCL ± P, and 236 with cleft palate only-NSCPO) and 693 unaffected Brazilian controls. Genomic ancestry was assessed with a set of 40 biallelic short insertion/deletion variants previously validated as ancestry informative markers of the Brazilian population. After adjustment of ancestry variations, allelic analysis revealed marginal associations between the CRISPLD2 rs4783099 T allele and increased risk for NSCPO (OR: 1.31, 95% CI: 1.05–1.62, P = 0.01) and between JARID2 rs2237138 and decreased NSCL ± P risk (OR: 0.80, 95% CI: 0.67–0.97, P = 0.02). Haplotype analysis indicated a lack of association between JARID2 haplotypes and non-syndromic oral cleft risk. Our results suggest that CRISPLD2 rs4783099 may represent a risk factor for NSCPO while JARID2 rs2237138 shows a protective effect against NSCL ± P in the Brazilian population.463232239CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESSem informaçãoSem informaçã

Genetic risk factors for nonsyndromic cleft lip with or without cleft palate in a brazilian population with high african ancestry

Nonsyndromic cleft lip with or without cleft palate (NSCL +/- P) is the most common orofacial birth defect, exhibiting variable prevalence around the world, often attributed to ethnic and environmental differences. Linkage analyses and genome-wide association studies have identified several genomic susceptibility regions for NSCL +/- P, mostly in European-derived or Asian populations. Genetic predisposition to NSCL +/- P is ethnicity-dependent, and the genetic basis of susceptibility to NSCL +/- P likely varies among populations. The population of Brazil is highly admixed, with highly variable ancestry; thus, the genetic determinants of NSCL +/- P susceptibility may be quite different. This study tested association of 8 single-nucleotide polymorphisms (SNPs), previously identified by genome-wide studies in other populations, with NSCL +/- P in a Brazilian population with high African ancestry. SNPs rs560426, rs642961, rs1530300, rs987525, rs3758249, rs7078160, rs17085106, and rs13041247 were genotyped in 293 Brazilian patients with NSCL +/- P and 352 unaffected Brazilian controls. Each sample was also genotyped for 40 biallelic short insertion/deletion polymorphic markers to characterize genetic ancestry. The average African ancestry background was 31.1% for the NSCL +/- P group and 36.7% for the control group. After adjustment for ancestry and multiple testing, the minor alleles of rs3758249 (OR: 1.58, 95% CI: 1.25-2.01, P=0.0001) and rs7078160 (OR: 1.59, 95% CI: 1.21-2.07, P=0.0002) were significantly associated with risk of NSCL +/- P. Polymorphisms located in IRF6 (rs642961) and 8q24 (rs1530300 and rs987525) showed marginal associations in this Brazilian population with high African ancestry. These results indicate that rs3758249 at 9q22 and rs7078160 at 10q25.3 represent risk loci for NSCL +/- P in the Brazilian population with high African ancestry1671023442349CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DA BAHIA - FAPESBFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP470606/2011-8; 484292/2013-700007/014470606/2011-8; 484292/2013-

Association between genes involved in craniofacial development and nonsyndromic cleft lip and/or palate in the brazilian population

To determine the association of single-nucleotide polymorphisms (SNPs) in genes related to craniofacial development, which were previously identified as susceptibility signals for nonsyndromic oral clefts, in Brazilians with nonsyndromic cleft lip and/or palate (NSCL/P). The SNPs rs748044 (TNP1), rs1106514 (MSX1), rs28372960, rs15251 and rs2569062 (TCOF1), rs7829058 (FGFR1), rs1793949 (COL2A1), rs11653738 (WNT3), and rs242082 (TIMP3) were assessed in a family-based transmission disequilibrium test (TDT) and a structured case-control analysis based on the individual ancestry proportions. The SNPs were initially analyzed by TDT, and polymorphisms showing a trend toward excess transmission were subsequently studied in an independent case-control sample. The study sample consisted of 189 case-parent trios of nonsyndromic cleft lip with or without cleft palate (NSCL±P), 107 case-parent trios of nonsyndromic cleft palate (NSCP), 318 isolated samples of NSCL±P, 189 isolated samples of NSCP, and 599 healthy controls. Association of alleles with NSCL/P pathogenesis. Preferential transmission of SNPs rs28372960 and rs7829058 in NSCL±P trios and rs11653738 in NSCP trios (P = .04) were observed, although the structured case-control analysis did not confirm these associations. The haplotype T-C-C formed by TCOF1 SNPs rs28372960, rs15251, and rs2569062 was more frequently transmitted from healthy parents to NSCL±P offspring, but the P value (P = .01) did not withstand Bonferroni correction for multiple tests. With the modest associations, our results do not support the hypothesis that TNP1, MSX1, TCOF1, FGFR1, COL2A1, WNT3, and TIMP3 variants are risk factors for nonsyndromic oral clefts in the Brazilian population.535550556CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE MINAS GERAIS - FAPEMIGFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPSem informaçãoSem informaçãoSem informaçãoSem informaçã