Lnc2-aren karakterizazio molekularra giza pankreako beta zeluletan

Abstract

[EN] Type I diabetes (T1D) is an autoimmune disease in which due to the destruction of pancreatic beta cells, patients become insulin dependent for life. Both, environmental (e.g. viral infections) and genetic factors, contribute to T1D susceptibility. Accumulating scientific evidence support the implication of viral infections in T1D development. Thus, upon a viral infection in pancreatic beta cells, the immune system is activated, leading to pancreatic beta cell inflammation (insulitis). Inflammation leads to the activation of several pro-inflammatory and pro-apoptotic pathways in beta cells that ends in pancreatic beta cell destruction. In recent years, long non coding RNAs have been associated with T1D. Thus, the main objective of the present project was to characterize the function of a T1D-associated lncRNA named Lnc2 at the pancreatic beta cell level. To this aim, pancreatic beta cells (EndoC-H1) were exposed to pro-inflammatory cytokines or intracellular synthetic viral double stranded RNA (PIC) to create a pro-inflammatory environment and analyse the expression of Lnc2. Besides that, taking into account that Lnc2 overlaps with BTNL2 in the human genome, the effect of Lnc2 on BTNL2 expression was also studied. The results showed that both, pro-inflammatory cytokines and PIC, upregulate Lnc2 expression in beta cells. Moreover, these results revealed that Lnc2 participates in the regulation of BTNL2 expression at the pancreatic beta cell level. Finally, the results of this work suggested that Lnc2 might play a role in pancreatic beta cells apoptosis. In conclusion, Lnc2 could be involved in pancreatic beta cell death, probably through the regulation of a gene that plays a role in apoptosis. Furthermore, since Lnc2 overlaps with BTNL2 in the human genome and Lnc2 regulates BTNL2 expression, the effect of Lnc2 overexpression in beta cell apoptosis might be driven by BTNL2.

[EUS] 1 motako diabetesa (DM1; Diabetes Mellitus 1) gaixotasun autoimmune bat da non, pankreako beta zelulen suntsipenaren ondorioz gaixoak intsulina tratamenduarekiko dependente bihurtzen diren. DM1aren garapena faktore genetiko zein ingurune faktoreen (infekzio biralak, besteak beste) menpekoa da. Ebidentzia zientifiko ugari daude birusak eta DM1aren garapena erlazionatzen dituztenak. Hauen arabera, birus batek beta zelulak infektatzean, sistema immunitarioa aktibatzen da, pankreako irletan hantura eraginez (insulitisa). Hanturak pankreako beta zeluletan zenbait bidezidor pro-inflamatorio eta pro-apoptotiko aktibatuko lituzke, pankreako beta zelulen suntsipena eraginez. Azken urteetan, RNA luze ez kodetzaileak DM1arekin asoziatu dira eta, lan honetan, DM1arekin asoziatutako polimorfismoa duen lncRNA bat izan da aztergai, Lnc2 deritzona. Horregatik, Gradu Amaierako Lan honen helburua Lnc2-aren karakterizazio molekularra egitea izan da, giza pankreako beta zeluletan bete dezakeen funtzioa aztertzeko asmoz. Horretarako, pankreako beta zelulak (EndoC-H1) hantura sortzen duten zitokina eta harizpi bikoitzeko RNA birikoarekin (PIC) tratatu dira; beta zeluletan baldintza pro-inflamatorioak sortu eta, Lnc2-aren adierazpena aztertzeko. Horretaz gain, giza genoman Lnc2-arekin gainjarrita dagoen BTNL2 genea ere aztertu da, Lnc2-ak gene horren adierazpenean izan dezakeen eragina zehaztuz. Emaitzek adierazi dute Lnc2-aren adierazpena igo egiten dela beta zelulak zitokina zein PICarekin tratatzean. Gainera, Lnc2-ak nolabait BTNL2 genearen adierazpena erregulatzen duela ikusi da. Horretaz gain, Lnc2-ak DM1ean ematen den pankreako beta zelulen apoptosian parte hartzen duela ikusi da. Ondorioz, Lnc2-ak DM1ean pankreako beta zeluletan ematen den suntsipenean parte hartzen duela esan daiteke, apoptosian parte hartzen duen gene baten erregulazioaren bidez, adibidez. Gainera, genoman BTNL2arekin gainjarrita agertzen denez eta Lnc2-ak BTNL2aren adierazpena erregulatzen duenez, gerta daiteke Lnc2 geneak beta zelulen apoptosian duen efektua BTNL2 genearen bitartekoa izatea.