Characterization of a Novel Long Non-coding RNA Involved in Thyroid Differentiation

Thyroid is the endocrine gland that most frequently undergoes to congenital disorders or neoplastic transformation and despite its organogenesis is well characterized, molecular bases of early thyroid differentiation are still obscure. During last years, long non-coding RNAs (lncRNA) have acquired increasing relevance in many biological processes, such as differentiation and cancer. In E10.5 mouse thyroid bud the most enriched transcript resulted to be a poorly characterized lncRNA, that we named Thybe1 (thyroid bud enriched 1). Thybe1 is an antisense transcript of the protein-coding gene klhl14, also enriched in thyroid bud, to which it partially overlaps in a head-to-head arrangement. To shed light on its role, in this work we characterize such novel lncRNA, investigating its role in thyroid differentiation and its possible mechanism of action. Interestingly, our data reveal that Thybe1 is required for thyroid differentiation, in vitro, and is able to compete, for miR182a-5p binding, with Pax8 and Bcl2, both playing a key role in thyroid survival and differentiation. Moreover, we observed that Thybe1 is dramatically repressed during thyroid carcinogenesis, being inversely correlated with miR182a-5p expression, both in vitro and in vivo. Furthermore, we noted that Thybe1 expression positively correlates with that of Klhl14 in several cell types, suggesting that this lncRNA is also able to act in cis on this target. In conclusion we describe for the first time a lncRNA involved in thyroid differentiation and carcinogenesis, and start to highlight its ability to act as a competing endogenous RNA for key developmental genes, thus identifying a novel candidate gene playing a role in thyroid development defects and cancer

Modellazione del comportamento dinamico delle opere di sostegno flessibili

Leonardo Cascini, Renato Lancellotta, Cristina Jomm

Modulating iron spin states with radical ligands: a density functional theoretical study

The ground state electronic structures of [FeIIIX(LISQ)2]0 where X is a halide (F−, Cl−, Br−, I−) or pseudo‐halide (N3−, NCS−) and (LISQ)1− is the o ‐iminobenzosemiquinonato π‐radical ligand, have been calculated using DFT at the B3LYP* level of theory. The modified functional with 15% Hartree‐Fock exchange is required to successfully reproduce the spin ground state of the complex as either S =3/2 for X=F−, Cl− and NCS−, or S =1/2 for X=Br−, I− and N3−. The difference in ground state stems from an S Fe=5/2→S Fe=3/2 spin transition at the iron ion, prompted by the donor properties of the apical ligand. The computational methodology was validated through accurate calculation of the Mössbauer parameters. The redox chemistry of the o ‐aminophenolate ligand was examined for the putative five‐membered electron transfer series for [FeIIIF(LISQ)2]z and [FeIIII(LISQ)2]z (z =2+1+, 0, 1−, 2−). The redox chemistry is entirely ligand‐centered with retention of the ferric ion, where only the strong ligand field provided by a fully reduced o ‐anilinophenolate(2−) ligand in conjunction with a soft apical donor will support an intermediate‐spin Fe(III) central ion

Tissue- and Cell Type-Specific Expression of the Long Noncoding RNA Klhl14-AS in Mouse

lncRNAs are acquiring increasing relevance as regulators in a wide spectrum of biological processes. The extreme heterogeneity in the mechanisms of action of these molecules, however, makes them very difficult to study, especially regarding their molecular function. A novel lncRNA has been recently identified as the most enriched transcript in mouse developing thyroid. Due to its genomic localization antisense to the protein-encoding Klhl14 gene, we named it Klhl14-AS. In this paper, we highlight that mouse Klhl14-AS produces at least five splicing variants, some of which have not been previously described. Klhl14-AS is expressed with a peculiar pattern, characterized by diverse relative abundance of its isoforms in different mouse tissues. We examine the whole expression level of Klhl14-AS in a panel of adult mouse tissues, showing that it is expressed in the thyroid, lung, kidney, testis, ovary, brain, and spleen, although at different levels. In situ hybridization analysis reveals that, in the context of each organ, Klhl14-AS shows a cell type-specific expression. Interestingly, databases report a similar expression profile for human Klhl14-AS. Our observations suggest that this lncRNA could play cell type-specific roles in several organs and pave the way for functional characterization of this gene in appropriate biological contexts

A ceRNA circuitry involving the long noncoding RNA KLHL14-AS, PAX8, and BCL2 drives thyroid carcinogenesis

Klhl14-AS is a long noncoding RNA expressed since early specification of thyroid bud and is the most enriched gene in the mouse thyroid primordium at E10.5. Here, we studied its involvement in thyroid carcinogenesis by analyzing its expression in cancer tissues and different models of neoplastic transformation. Compared with normal thyroid tissue and cells, Klhl14-AS was significantly downregulated in human thyroid carcinoma tissue specimens, particularly the anaplastic histotype, thyroid cancer cell lines, and rodent models of thyroid cancer. Downregulating the expression of Klhl14-AS in normal thyroid cells decreased the expression of thyroid differentiation markers and cell death and increased cell viability. These effects were mediated by the binding of Klhl14-AS to two miRNAs, Mir182-5p and Mir20a-5p, which silenced Pax8 and Bcl2, both essential players of thyroid differentiation. MIR182-5p and MIR20a-5p were upregulated in human thyroid cancer and thyroid cancer experimental models and their effects on Pax8 and Bcl2 were rescued by Klhl14-AS overexpression, confirming Klhl14-AS as a ceRNA for both Pax8 and Bcl2. This work connects deregulation of differentiation with increased proliferation and survival in thyroid neoplastic cells and highlights a novel ceRNA circuitry involving key regulators of thyroid physiology. Significance: This study describes a new ceRNA with potential tumor suppression activity and helps us better understand the regulatory mechanisms during thyroid differentiation and carcinogenesis

Outcomes of pregnancies after kidney transplantation: lessons learned from CKD. A comparison of transplanted, nontransplanted chronic kidney disease patients and low-risk pregnancies: a multicenter nationwide analysis.

BACKGROUND: Kidney transplantation (KT) may restore fertility in CKD. The reasons why materno-foetal outcomes are still inferior to the overall population are only partially known. Comparison with the CKD population may offer some useful insights for management and counselling.Aim of this study was to analyse the outcomes of pregnancy after KT, compared with a large population of non-transplanted CKD patients and with low-risk control pregnancies, observed in Italy the new millennium. METHODS: We selected 121 live-born singletons after KT (Italian study group of kidney in pregnancy, national coverage about 75%), 610 live-born singletons in CKD and 1418 low-risk controls recruited in 2 large Italian Units, in the same period (2000-2014). The following outcomes were considered: maternal and foetal death; malformations; preterm delivery; small for gestational age baby (SGA); need for the neonatal intensive care unit (NICU); doubling of serum creatinine or increase in CKD stage. Data were analysed according to kidney diseases, renal function (staging according to CKD-EPI), hypertension, maternal age, partity, ethnicity. RESULTS: Materno-foetal outcomes are less favourable in CKD and KT as compared with the low-risk population. CKD stage and hypertension are important determinants of results. KT patients with e-GFR >90 have worse outcomes compared with CKD stage 1 patients; the differences level off when only CKD patients affected by glomerulonephritis or systemic diseases ('progressive CKD') are compared with KT. In the multivariate analysis, risk for preterm and early-preterm delivery was linked to CKD stage (2-5 versus 1: RR 3.42 and 3.78) and hypertension (RR 3.68 and 3.16) while no difference was associated with being a KT or a CKD patient. CONCLUSIONS: The materno-foetal outcomes in patients with kidney transplantation are comparable with those of nontransplanted CKD patients with similar levels of kidney function impairment and progressive and/or immunologic kidney diseas