Publications
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Heinicke, Fatima; Zhong, Xiangfu; Flåm, Siri Tennebø; Breidenbach, Johannes; Leithaug, Magnus & Mæhlen, Marthe Thoresen
[Show all 14 contributors for this article]
(2021).
MicroRNA Expression Differences in Blood-Derived CD19+ B Cells of Methotrexate Treated Rheumatoid Arthritis Patients.
Frontiers in Immunology.
ISSN 1664-3224.
12.
doi:
10.3389/fimmu.2021.663736.
Full text in Research Archive
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Rheumatoid arthritis (RA) is a complex disease with a wide range of underlying susceptibility factors. Recently, dysregulation of microRNAs (miRNAs) in RA have been reported in several immune cell types from blood. However, B cells have not been studied in detail yet. Given the autoimmune nature of RA with the presence of autoantibodies, CD19+ B cells are a key cell type in RA pathogenesis and alterations in CD19+ B cell subpopulations have been observed in patient blood. Therefore, we aimed to reveal the global miRNA repertoire and to analyze miRNA expression profile differences in homogenous RA patient phenotypes in blood-derived CD19+ B cells. Small RNA sequencing was performed on CD19+ B cells of newly diagnosed untreated RA patients (n=10), successfully methotrexate (MTX) treated RA patients in remission (MTX treated RA patients, n=18) and healthy controls (n=9). The majority of miRNAs was detected across all phenotypes. However, significant expression differences between MTX treated RA patients and controls were observed for 27 miRNAs, while no significant differences were seen between the newly diagnosed patients and controls. Several of the differentially expressed miRNAs were previously found to be dysregulated in RA including miR-223-3p, miR-486-3p and miR-23a-3p. MiRNA target enrichment analysis, using the differentially expressed miRNAs and miRNA-target interactions from miRTarBase as input, revealed enriched target genes known to play important roles in B cell activation, differentiation and B cell receptor signaling, such as STAT3, PRDM1 and PTEN. Interestingly, many of those genes showed a high degree of correlated expression in CD19+ B cells in contrast to other immune cell types. Our results suggest important regulatory functions of miRNAs in blood-derived CD19+ B cells of MTX treated RA patients and motivate for future studies investigating the interactive mechanisms between miRNA and gene targets, as well as the possible predictive power of miRNAs for RA treatment response.
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Heinicke, Fatima; Zhong, Xiangfu; Zucknick, Karola Manuela; Breidenbach, Johannes; Sundaram, Arvind Yegambaram Meenakshi & Flåm, Siri Tennebø
[Show all 11 contributors for this article]
(2020).
An extension to: Systematic assessment of commercially available low-input miRNA library preparation kits.
RNA Biology.
ISSN 1547-6286.
17(9),
p. 1284–1292.
doi:
10.1080/15476286.2020.1761081.
Full text in Research Archive
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High-throughput sequencing has emerged as the favoured method to study microRNA (miRNA) expression, but biases introduced during library preparation have been reported. We recently compared the performance (sensitivity, reliability, titration response and differential expression) of six commercially-available kits on synthetic miRNAs and human RNA, where library preparation was performed by the vendors. We hereby supplement this study with data from two further commonly used kits (NEBNext, NEXTflex) whose manufacturers initially declined to participate. NEXTflex demonstrated the highest sensitivity, which may reflect its use of partially-randomized adapter sequences, but overall performance was lower than the QIAseq and TailorMix kits. NEBNext showed intermediate performance. We reaffirm that biases are kit specific, complicating the comparison of miRNA datasets generated using different kits.
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Zhong, Xiangfu; Heinicke, Fatima & Rayner, Simon
(2019).
miRBaseMiner, a tool for investigating miRBase content.
RNA Biology.
ISSN 1547-6286.
16(11),
p. 1534–1546.
doi:
10.1080/15476286.2019.1637680.
Full text in Research Archive
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microRNAs are small non-coding RNA molecules playing a central role in gene regulation. miRBase is the
standard reference source for analysis and interpretation of experimental studies. However, the richness
and complexity of the annotation is often underappreciated by users. Moreover, even for experienced
users, the size of the resource can make it difficult to explore annotation to determine features such as
species coverage, the impact of specific characteristics and changes between successive releases.
A further consideration is that each new miRBase release contains entries that have had limited review
and which may subsequently be removed in a future release to ensure the quality of annotation. To aid
the miRBase user, we developed a software tool, miRBaseMiner, for investigating miRBase annotation
and generating custom annotation sets. We apply the tool to characterize each release from v9.2 to v22
to examine how annotation has changed across releases and highlight some of the annotation features
that users should keep in mind when using for miRBase for data analysis. These include: (1) entries with
identical or very similar sequences; (2) entries with multiple annotated genome locations; (3) hairpin
precursor entries with extremely low-estimated minimum free energy; (4) entries possessing reverse
complementary; (5) entries with 3ʹ poly(A) ends. As each of these factors can impact the identification of
dysregulated features and subsequent clinical or biological conclusions, miRBaseMiner is a valuable
resource for any user using miRBase as a reference source.
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Heinicke, Fatima; Zhong, Xiangfu; Zucknick, Manuela; Breidenbach, Johannes; Sundaram, Arvind Yegambaram Meenakshi & Flåm, Siri Tennebø
[Show all 21 contributors for this article]
(2019).
Systematic assessment of commercially available low-input miRNA library preparation kits.
RNA Biology.
ISSN 1547-6286.
17(1),
p. 75–86.
doi:
10.1080/15476286.2019.1667741.
Full text in Research Archive
Show summary
High-throughput sequencing is increasingly favoured to assay the presence and abundance of microRNAs (miRNAs) in biological samples, even from low RNA amounts, and a number of commercial vendors now offer kits that allow miRNA sequencing from sub-nanogram (ng) inputs. Although biases introduced during library preparation have been documented, the relative performance of current reagent kits has not been investigated in detail. Here, six commercial kits capable of handling <100ng total RNA input were used for library preparation, performed by kit manufactures, on synthetic miRNAs of known quantities and human total RNA samples. We compared the performance of miRNA detection sensitivity, reliability, titration response and the ability to detect differentially expressed miRNAs. In addition, we assessed the use of unique molecular identifiers (UMI) sequence tags in one kit. We observed differences in detection sensitivity and ability to identify differentially expressed miRNAs between the kits, but none were able to detect the full repertoire of synthetic miRNAs. The reliability within the replicates of all kits was good, while larger differences were observed between the kits, although none could accurately quantify the relative levels of the majority of miRNAs. UMI tags, at least within the input ranges tested, offered little advantage to improve data utility. In conclusion, biases in miRNA abundance are heavily influenced by the kit used for library preparation, suggesting that comparisons of datasets prepared by different procedures should be made with caution. This article is intended to assist researchers select the most appropriate kit for their experimental conditions.
View all works in Cristin
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Heinicke, Fatima; Rieder, Anne; Grimmer, Stine; Samuelsen, Anne Berit C. & Knutsen, Svein Halvor
(2015).
Establishing a gene expression system to screen the effect of dietary fibers and their metabolites on selected aspects of colon cancer development.
7Letras.
View all works in Cristin
Published
Nov. 11, 2021 12:26 PM
- Last modified
Jan. 10, 2023 3:32 PM