Combined molecular and mathematical analysis of long noncoding RNAs expression in fine needle aspiration biopsies as novel tool for early diagnosis of thyroid cancer

Possieri, Corrado; Locantore, Pietro; Salis, Chiara; Bacci, Lorenza; Aiello, Aurora; Fadda, Guido; Crea, Carmela De; Raffaelli, Marco; Bellantone, Rocco Domenico Alfonso; Grassi, Claudio; Strigari, Lidia; Farsetti, Antonella; Pontecorvi, Alfredo; Nanni, Simona

doi:10.1007/s12020-020-02508-w

Cited by 19 publications

(20 citation statements)

References 31 publications

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“…The performance of a smaller mutation panel combined with an miRNA risk classifier (ThyGeNEXT + ThyraMIR) was recently validated in a multicenter study [89], which reported good sensitivity (95%) and specificity (90%) for malignancy in indeterminate nodules. Some other, potentially less expensive, molecular tests were recently proposed, but need prospective and multicenter validation; these include a dual-component molecular assay (an extended DNA and RNA mutation panel plus a single miRNA expression level) [90], and the analysis of selected long non-coding RNAs (MALAT1, HOTAIR, PVT1) [91].…”

Section: Molecular Diagnosticsmentioning

confidence: 99%

Thyroid Nodule Characterization: How to Assess the Malignancy Risk. Update of the Literature

et al. 2021

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Ultrasound (US) is the first imaging modality for thyroid parenchyma evaluation. In the last decades, the role of ultrasound has been improved with the introduction of new US software, such as contrast-enhanced ultrasound (CEUS) and US-elastography (USE). USE is nowadays recognized as an essential part of the multiparametric ultrasound (MPUS) examination, in particular for the indeterminate thyroid nodule with possible fine-needle aspiration cytology (FNAC) number reduction; even if further and larger studies are needed to validate it. More controversial is the role of CEUS in thyroid evaluation, due to its high variability in sensitivity and specificity. Semi-automatic US systems based on the computer-aided diagnosis (CAD) system are producing interesting results, especially as an aid to less experienced operators. New knowledge on the molecular mechanisms involved in thyroid cancer is allowing practitioners to identify new genomic thyroid markers that could reduce the number of “diagnostic” thyroidectomies. We have therefore drawn up an updated representation of the current evidence in the literature for thyroid nodule multiparametric ultrasound (MPUS) evaluation with particular regard to USE, the US CAD system and CEUS.

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Section: Molecular Diagnosticsmentioning

confidence: 99%

Thyroid Nodule Characterization: How to Assess the Malignancy Risk. Update of the Literature

et al. 2021

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“…We first sought to profile the expression of H1R and H2R in MSNs of the NAc by using a single cell approach. For this, we assessed H2R and H1R mRNA levels in MSNs using a protocol that combines whole‐cell patch‐clamp recordings with high‐quality single cell RNA analysis by ddPCR (single‐cell qRT‐ddPCR) (Cadwell et al., 2016; Possieri et al., 2021). As shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…RNA was extracted from mouse NAc and cortex (Ctx) tissues using Trizol after tissue homogenization (homogenizer VDI12, VWR). RNA extraction, cDNA preparation and real-time PCR with Syber Green on QuantStudio 5 System (Applied Biosystems, Foster City, CA, USA) was performed as in Possieri et al (2021). Primers are as follows: HRH1 F 5 -CATCACTCCCTAGGACAGCC-3 , R 5 -GTCCTGTT CCCCTCACACAT-3 ; HRH2 F 5 -AGCTCCTATGACC CCAGAAAGAGT-3 , R 5 -TGGTGCCTGTTTCCATCG A-3 ; KCND2 F 5 -GGCAGCCTTCTGGTACACCAT-3 ; R 5 -GCACGGGTAGCGCAATG-3 ; GAPDH F 5 -TGC ACCACCACCTGCTTAGC-3 , R 5 -GTCTTCTGGGTG GCAGTGATG-3 .…”

Section: Rna Extraction and Rt-pcrmentioning

confidence: 99%

Activation of histamine type 2 receptors enhances intrinsic excitability of medium spiny neurons in the nucleus accumbens

Aceto

Nardella

Nanni

et al. 2022

The Journal of Physiology

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Histaminergic neurons are exclusively located in the hypothalamic tuberomammillary nucleus, from where they project to many brain areas including the nucleus accumbens (NAc), a brain area that integrates diverse monoaminergic inputs to coordinate motivated behaviours. While the NAc expresses various histamine receptor subtypes, the mechanisms by which histamine modulates NAc activity are still poorly understood. Using whole‐cell patch‐clamp recordings, we found that pharmacological activation of histamine 2 (H2) receptors elevates the excitability of NAc medium spiny neurons (MSNs), while activation of H1 receptors failed to significantly affect MSN excitability. The evoked firing of MSNs increased after seconds of local H2 agonist administration and remained elevated for minutes. H2 receptor (H2R) activation accelerated subthreshold depolarization in response to current injection, reduced the latency to fire, diminished action potential afterhyperpolarization and increased the action potential half‐width. The increased excitability was protein kinase A‐dependent and associated with decreased A‐type K+ currents. In addition, selective pharmacological inhibition of the Kv4.2 channel, the main molecular determinant of A‐type K+ currents in MSNs, mimicked and occluded the increased excitability induced by H2R activation. Our results indicate that histaminergic transmission in the NAc increases MSN intrinsic excitability through H2R‐dependent modulation of Kv4.2 channels. Activation of H2R will significantly alter spike firing in MSNs in vivo, and this effect could be an important mechanism by which these receptors mediate certain aspects of goal‐induced behaviours. Key points Histamine is synthesized and released by hypothalamic neurons of the tuberomammillary nucleus and serves as a general modulator for whole‐brain activity including the nucleus accumbens. Histamine receptors type 2 (HR2), which are expressed in the nucleus accumbens, couple to Gαs/off proteins which elevate cyclic adenosine monophosphate levels and activate protein kinase A. Whole‐cell patch‐clamp recordings revealed that H2R activation increased the evoked firing in medium spiny neurons of the nucleus accumbens via protein kinase A‐dependent mechanisms. HR2 activation accelerated subthreshold depolarization in response to current injection, reduced the latency to fire, diminished action potential medium after‐hyperpolarization and increased the action potential half‐width. HR2 activation also reduced A‐type potassium current. Selective pharmacological inhibition of the Kv4.2 channel mimicked and occluded the increased excitability induced by H2R activation.

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“…Besides their involvement in genomic imprinting, inactivation of chromosome X, maintenance of pluripotency, and the formation of different organs via changes in chromatin, transcription, and translation, they are also known to act as tumor suppressor genes or oncogenes. Recently, Possieri et al (37) analyzed six cancer-associated lncRNAs (MALAT1, NEAT1, HOTAIR, H19, PVT1, MEG3) in 135 FNA samples, with MALAT1, PVT1, and HOTAIR showing a significant differentiation capability between malignant and benign nodules (P < 0.0001).…”

Section: Contribution Of Epigenetic To Cytology Diagnostic Challengesmentioning

confidence: 99%

Translational Potential of Epigenetic-Based Markers on Fine-Needle Aspiration Thyroid Specimens

et al. 2021

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The awareness of epigenetic alterations leading to neoplasia attracted the attention of researchers toward its potential use in the management of cancer, from diagnosis to prognosis and prediction of response to therapies. Our group has focused its attention on the epigenomics of thyroid neoplasms. Although most of the epigenetic studies have been applied on histological samples, the fact is that cytology, through fine-needle aspiration, is a primary diagnostic method for many pathologies, of which thyroid nodules are one of the most paradigmatic examples. This has led to an increasing literature report of epigenetic studies using these biological samples over the past decade. In this review, our group aimed to document recent research of epigenetic alterations and its associated assessment techniques, based on cytology material. Our review covers the main epigenetic categories—DNA methylation, histone modification, and RNA-silencing—whose evidence in thyroid cytology samples may represent solid soil for future prospectively designed studies aiming at validating patterns of epigenetic alterations and their potential use in the clinical management of thyroid neoplasms.

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Combined molecular and mathematical analysis of long noncoding RNAs expression in fine needle aspiration biopsies as novel tool for early diagnosis of thyroid cancer

Cited by 19 publications

References 31 publications

Thyroid Nodule Characterization: How to Assess the Malignancy Risk. Update of the Literature

Thyroid Nodule Characterization: How to Assess the Malignancy Risk. Update of the Literature

Activation of histamine type 2 receptors enhances intrinsic excitability of medium spiny neurons in the nucleus accumbens

Translational Potential of Epigenetic-Based Markers on Fine-Needle Aspiration Thyroid Specimens

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