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Novel 3D-printed hollow microneedles facilitate safe, reliable, and informative sampling of perilymph from guinea pigs

ABSTRACT: Inner ear diagnostics is limited by the inability to atraumatically obtain samples of inner ear fluid. The round window membrane (RWM) is an attractive portal for accessing perilymph samples as it has been shown to heal within one week after the introduction of microperforations. A 1 µL volume of perilymph is adequate for proteome analysis, yet the total volume of perilymph within the scala tympani of the guinea pig is limited to less than 5 µL. This study investigates the safety and reliability of a novel hollow microneedle device to aspirate perilymph samples adequate for proteomic analysis.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Cavia (ncbitaxon:10140)

SUBMITTER: Jeffrey W.Kysar Anil K. Lalwani Lewis M. Brown

PROVIDER: MSV000086817 | MassIVE | Mon Feb 08 10:27:00 GMT 2021

REPOSITORIES: MassIVE

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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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Novel 3D-printed hollow microneedles facilitate safe, reliable, and informative sampling of perilymph from guinea pigs

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