Submitted by Pós Imunologia (ppgimicsufba@gmail.com) on 2017-02-07T18:12:52Z No. of bitstreams: 1 Dissertação Gerson 2016.pdf: 3140570 bytes, checksum: f563f94e781752cf57592ebd916b6bc8 (MD5) / Approved for entry into archive by Delba Rosa (delba@ufba.br) on 2017-02-08T16:09:34Z (GMT) No. of bitstreams: 1 Dissertação Gerson 2016.pdf: 3140570 bytes, checksum: f563f94e781752cf57592ebd916b6bc8 (MD5) / Made available in DSpace on 2017-02-08T16:09:34Z (GMT). No. of bitstreams: 1 Dissertação Gerson 2016.pdf: 3140570 bytes, checksum: f563f94e781752cf57592ebd916b6bc8 (MD5) / CAPES / Asma e atopia são condições determinadas por fatores ambientais e genéticos Vários estudos de associação do genoma têm sido realizados para tentar entender os componentes genéticos de tais condições. Os genes IL33 e IL1RL1 são os mais replicados em estudos do tipo GWAS em todo o mundo. A citocina IL-33 e o seu receptor ligado à membrana (ST2L) ou sua forma solúvel (sST2), em conjunto são potentes moduladores de inflamação do tipo Th2. Quando ligada ao ST2L, IL-33 produzida por múltiplas células da resposta inata e adaptativa, induz citocinas pró-inflamatórias, tais como IL-4, IL-5 e IL-13, aumentando a resposta e inflamação Th2, principal característica da asma e alergias. Por outro lado, quando a IL-33 se liga ao sST2, neutraliza o seu efeito, impedindo sua ligação ao ST2L. Vários polimorfismos nestes genes têm sido associados com a asma e atopia. Assim, os fatores genéticos que afetam IL33 e IL1RL1 podem influenciar a susceptibilidade para asma e atopia. Neste contexto, este estudo teve como objetivo avaliar a influência de polimorfismos em IL33 e IL1RL1 na asma e atopia em uma população Latina. Estratégias diferentes foram usadas para, um estudo de coorte de asma leve e um estudo caso-controle de asma grave. O alelo A para rs1041973 em IL1RL1 na coorte SCAALA foi positivamente associado com IL-5 produção (OR 1,36, IC 95% 1,09-1,84, P=0,044) IgE específica (OR 1,40, IC 95% 1,07-1,84, P=0,013) e SPT (OR 1,48, IC 95% 1,08-2,03, P=0,014), ambos contra o ácaro B. tropicalis. Além disso, indivíduos atópicos com o genótipo AA de rs1041973 mostraram uma diminuição da produção de sST2 comparado com indivíduos com os genótipos AC e CC (P <0,05). O alelo G do SNP IL33 rs12551256 foi negativamente associado com asma (OR 0,71, IC 95% 0,53-0,94, P=0,017). Em relação ao estudo caso controle do ProAR, o alelo A do rs1420101 em IL1RL1, foi positivamente associado com asma atópica (OR 1,29, IC 95% 1,05-1,66, P=0,046) e negativamente com FEV1 (BETA -2,37, IC 95% -4,67; -0,07, P=0,043). Além disso, este mesmo alelo mostrou uma diferença estatisticamente significate com uma menor produção de sST2 plasmático em indivíduos controle (P <0,001). O alelo C do rs2381416 foi positivamente associado com SPT para A. flavus. (OR 7,2, IC 95% 1,05-3,37, P=0,033), epitélio de cão (OR 1,52, IC 95% 1,19-3,47, P=0,009) e gato (OR 1,52, IC 95% 1,04-2,63, P=0,048). Estes dados sugerem que os SNPs nos genes IL33 e IL1RL1 podem ter um impacto sobre o desenvolvimento de asma e alergia na população brasileira. No entanto, mais estudos devem ser realizados para elucidar o impacto funcional de tais polimorfismos aqui descritos no desenvolvimento de asma e atopia. / QUEIROZ, Gerson de Almeida. STUDY OF INFLUENCE OF IL33 AND IL1RL1 POLYMORPHISMS IN SEVERE ASTHMA. 93f. 2016. Dissertação (Mestrado) - Instituto de Ciências da Saúde, Universidade Federal da Bahia. Asthma and atopy are conditions determined by environmental and genetic factors. Several genome-wide association studies have been conducted to try to understand the genetic components of such conditions. The IL33 and IL1RL1 are the most replicated genes in GWAS studies worldwide. The cytokine IL-33 and its receptor, membrane bound (ST2L) or its soluble form (sST2) together are potent Th2-type inflammation modulators. When bound to ST2L, IL-33 produced by multiple cells of the innate and adaptive response, induce proinflammatory cytokines such as IL-4, IL-5 and IL-13, increasing the response and Th2 inflammation main feature of asthma and allergies. On the other hand, when IL-33 binds to sST2, neutralizes their effect by preventing its binding to ST2L. Several polymorphisms in these genes have been associated with asthma and atopy. Thus, genetic factors that affect IL33 and IL1RL1 may influence susceptibility to asthma and atopy. In this context, this study aimed to evaluate the influence of polymorphisms in IL33 and IL1RL1 in asthma and atopy in a Latino population. To verify that he have used to different strategies, a cohort study for mild asthma and a case-control study for severe asthma. The A allele for rs1041973 in IL1RL1 in SCAALA cohort was positively associated with IL-5 production (1.36 OR, 95% CI 1.09-1.84, p=0.044) specific IgE (OR 1.40, 95% CI 1.07-1.84, p=0.013) and SPT (OR 1.48, 1.08-2.03 95% CI, p=0.014), both against B. tropicalis mite. Furthermore, atopic individuals with the AA genotype of rs1041973 showed a decreased production of sST2 as compared to individuals with the AC and CC genotypes (P<0.05). The G allele of IL33 SNP was negatively associated with asthma (OR 0.71, 95% CI 0.53-0.94, p=0.017). Regarding the ProAR case control study the A allele of rs1420101 in IL1RL1 was positively associated with atopic asthma (OR 1.29, 95% CI 1.05-1.66, P=0.046) and negatively associated with FEV1 (BETA -2.37, 95% CI -4.67 ; -0.07, P=0.043). In addition, this same allele showed a statistically significant difference with a lower production of plasma sST2 in control subjects (P<0.001). The C allele of rs2381416 was positively associated with SPT to A. flavus. (OR 7.2, 95% CI 1.05 to 3.37, P = 0.033), dog (OR 1:52, 1:19 to 3:47 95%, P = 0.009) and cat epithelium (OR 1:52, 95% CI 1.04-2.63, P = 0.048). These data suggest that SNPs in IL33 and IL1RL1 genes may have an impact on the development of asthma and allergy in Brazilian population. However, further studies should be conducted to further elucidate the functional impact of such polymorphisms described herein in the development of asthma and atopy.

La leucémie aiguë à basophile du Nourrisson est un sous-type rare de leucémie aiguë myéloïde.Notre équipe avait précédemment participé à la caractérisation moléculaire de la translocationrécurrente t(X;6)(p11;q23) générant un gène de fusion MYB-GATA1 chez les nourrissons desexe masculin. Pour mieux comprendre son rôle, le facteur de transcription MYB-GATA1résultant de cette fusion a été exprimé dans des cellules progénitrices de l’hématopoïèsehumaine, CD34+ avant xénogreffe chez des souris immunodéficientes. Les cellules exprimantMYB-GATA1 présentaient une augmentation de l’expression des marqueurs d’immaturité(CD34), des marqueurs de la lignée granuleuse (CD33, CD117) et des signes de différenciationbasophile (CD203c, FcƐRI). Des cellules de lignée UT-7 ont également montré descaractéristiques de différenciation basophile après transduction par MYB-GATA1. Une analysetranscriptomique a permis de mettre en évidence 9 gènes dérégulés à la fois par la présence deMYB-GATA1 et par la différenciation basophile. L’augmentation de l’expression de 3 de cesgènes (CCL23, IL1RL1 et NTRK1) a été confirmée en RT-PCRq dans des cellules CD34+transduites avec MYB-GATA1. L’IL-33 (Interleukine 33) et le NGF (Nerve Growth Factor),les ligands respectifs de IL1RL1 et NTRK1, augmentent la différenciation basophile de cellulesUT-7 exprimant MYB-GATA1, démontrant l’importance de ces voies de signalisation dans ladifférenciation basophiles de cellules leucémiques et de cellules primaires de l’hématopoïèsehumaine CD34+. Enfin une expérience utilisant la luciférase a confirmé que MYB et MYBGATA1augmentaient l’activité des facteurs de transcription NTRK1 et IL1RL1 conduisant àl’acquisition de caractéristiques basophiles. Nos résultats soulignent ainsi l’importance desrécepteurs à l’IL-33 et au NGF dans la différenciation basophile des cellules normales etleucémiques. / Acute basophilic leukaemia (ABL) is a rare subtype of acute myeloblastic leukaemia. Wepreviously described a recurrent t(X;6)(p11;q23) translocation generating a MYB-GATA1fusion gene in male infants with ABL. To better understand its role, the chimeric MYB-GATA1transcription factor was expressed in CD34-positive hematopoietic progenitors which weretransplanted into immunodeficient mice. Cells expressing MYB-GATA1 showed increasedexpression of markers of immaturity (CD34), of granulocytic lineage (CD33, CD117) and ofbasophilic differentiation (CD203c, FcƐRI). UT-7 cells also showed basophilic differentiationafter MYB-GATA1 transfection. A transcriptomic study identified 9 genes deregulated by bothMYB-GATA1 and by basophilic differentiation. Induction of three of these genes (CCL23,IL1RL1 and NTRK1) was confirmed in MYB-GATA1-expressing CD34-positive cells byRTqPCR. IL-33 and NGF (Nerve Growth Factor), the ligands of IL1RL1 and NTRK1,respectively, enhanced the basophilic differentiation of MYB-GATA1-expressing UT-7 cells,thus demonstrating the importance of this pathway in basophilic differentiation of leukemiccells and CD34 positive primary cells. Finally, gene reporter assays confirmed that MYB andMYB-GATA1 activated NTRK1 and IL1RL1 transcription leading to basophilic skewing ofthe blasts. Our results highlight the role of IL-33 and NGF receptors in basophilic differentiationof normal and leukemic cells.