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Circulating non-coding RNAs in Huntington disease

ABSTRACT: We performed a study of whole non-coding RNA expression profiles, by microarray, in plasma from 9 patients with Huntington disease (CH) in early stage and 13 controls: 8 healthy subjects (HD) and 5 patients with schizophrenia (SC). The CH patients were under antipsychotic drugs, the most effective treatment option also in SC patient (we included in the study SC control group to minimize the impact of neuroleptic drugson the expression of non-coding RNAs and to identify a HD-specific set of non-coding RNAs). Blood samples were obtained by venous punctures in EDTA-tubes for plasma preparation. Cell- and platelet-free plasma was prepared following a step centrifugation protocol: samples were centrifuged at 1.500 g for 15′ at 4 °C and supernatant will be stored at −80 °C. RNA was extracted using Plasma/Serum Circulating RNA purification Kit (NORGEN) following the manufacturer’s instructions. Labeled RNA from each sample was analyzed by GeneChipmiRNA 2.0 Array (Affymetrix) according to the manufacturer’s instructions. Raw data were analyzed using Partek Genomic Suite software. Looking for differently expressed non-coding RNAs, we found a candidate HC-specific set compared to both control groups (HD and SC) (p<0.05 and │foldchanges│>1.5): hsa-miR-98 has proved to be down-regulated, while hsa-miR-323b-3p was up-regulated; 2 novel snoRNAs were up-regulated.

ORGANISM(S): synthetic construct Homo sapiens

PROVIDER: GSE167630 | GEO | 2021/05/03

REPOSITORIES: GEO

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Project description:Background Micro RNAs (miRNAs) are a class of small non-coding RNAs that are involved in post transcriptional gene expression regulation. miRNAS can be found in various bodily fluids including plasma. Recently circulating plasma miRNAs have been studied as biomarkers in various tumors including pituitary neuroendocrine tumors (PitNETs). However the identification of PitNET derived miRNAs in plasma remains a challenging task due to tumor volume, mass and miRNA dilution within blood. To our knowledge this is the first study that has used the NGS approach to characterize circulating miRNAs in plasma acquired from Bilateral petrosal sinus sampling (BIPSS) - a gold standard in diagnosis of ACTH secreting PitNETs. Results We sequenced plasma samples that were acquired from sinistral and dextral sides of sinus petrosus inferior and complementary plasma from peripheral venous blood (PVB). BIPSS samples were collected in three separate time periods: before administration of corticotropin releasing hormone (CRH), 5 minutes after stimulation and 15 minutes after stimulation with CRH. The highest amount of differentially expressed miRNAs was observed 5 minutes after CRH stimulation (20 upregulated, 14 downregulated). During the BIPSS procedure the highest amount of ACTH was released in the sinistral side during the 5th minute after the stimulation by CRH. In the plasma of sinistral side at the 5th minute we identified two miRNAs: hsa-miR-7-5p and hsa-miR-375-3p that were highly upregulated compared to other BIPSS samples and samples from PVB. Using clustering analysis we were able to distinguish plasma samples acquired from BIPSS from PVB plasma, indicating that the miRNA fraction characterized in this study has potentially PitNET borne origin. Conclusions In this study we provided the first insight into the landscape of circulating miRNAs in close proximity to the ACTH secreting PitNET. The data indicates that ACTH secreting PitNET releases two circulating miRNAs upon stimulation with CRH (hsa-mir-7-5p, hsa-mir-375-3p) alongside with ACTH implying the further studies of these miRNA as diagnostic markers.

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Circulating non-coding RNAs in Huntington disease

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