MicroRNA-146a induction during influenza H3N2 virus infection targets and regulates TRAF6 levels in human nasal epithelial cells (hNECs)
Authors: De-Yun Wang
Ze-Zhang Tao
Jing Liu
Vincent T Chow
Yuqin Deng
Kai Sen Tan
Yan Yan
Journal: Experimental Cell Research (2017)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: Human nasal epithelial cells (hNECs); Influenza H3N2 virus; Innate immunity; MicroRNA-146a-5p; TNF receptor associated factor-6 (TRAF6).
Affiliations: Department of Otolaryngology, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, PR China; Department of Otolaryngology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore., Department of Otolaryngology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore., Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore., Department of Otolaryngology, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, PR China. Electronic address: ., Department of Otolaryngology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore. Electronic address: .

Abstract:  We have previously shown that human nasal epithelial cells (hNECs) are highly permissive cells for respiratory viruses including influenza A virus (IAV) and respiratory syncytial virus. Recent studies have indicated that microRNAs (miRNAs) are involved in virus-host relationship, and this led us to investigate its essential roles in the in vitro hNECs model derived from multiple donors. By comparing the differential expression of miRNAs upon IAV infection among animal and cell line studies, candidates were selected with focus on the initial immune response. After infection of influenza H3N2 virus, hNECs showed constant increase virus titer at 24-72h post-infection (hpi); accompanied with a significantly elevated level of miR-146a-5p at 72 hpi. The exponential elevation of progeny virus titer correlated with a key influenza sensing Toll-like receptor (TLR)7 pathway. TLR7 downstream gene transcripts, myeloid differentiation primary response gene 88 (MyD88), interferon regulator factor 7 (IRF7), and interferon-β (IFN-β) were significantly upregulated at 48 and 72 hpi, while interleukin-1 receptor-associated kinase 1 (IRAK1) and TNF receptor associated factor-6 (TRAF6) were unchanged. Interestingly, when miR-146a was overexpressed with miRNA mimics prior to H3N2 infection, further decreased transcripts of TRAF6, but not IRAK1, were detected. By using the in vitro hNEC model, we demonstrated that H3N2-induced miR-146a specifically targets and regulates TRAF6 expression; but not IRAK expression in the nasal epithelium. We also found that unlike the cell model studies that lead to our studies, when ran across a heterogeneous model of different individual, miRNA signals were highly varied and the expression of most miRNAs, including miR-146a-5p, was more subdued compared to homogenous cell line model, highlighting a need for a more thorough analysis of miRNA signals and targets in a model more mimicking a clinical influenza infection. 

Retracted: No

PubPeer comments:
 Retracted [November 2017] after notification by the editors of PLoS of overlaps with [Zhao et al (2017)] . ---------------------------------------- *Asian Pacific Journal of Cancer Prevention (APJCP)* has retracted the article titled u201cResveratrol Inhibits Oesophageal Adenocarcinoma Cell Proliferation via AMP-activated Protein Kinase Signalingu201d(1) for reason of having duplicated contents brought to the attention of APJCPu2019s editorial office by the following email content: u201cDear Editors of *Gastroenterology and Hepatology*, *Acta Pharmacologica Sinica*, *Asian Pac J Cancer Prev*, *Clinical and Experimental Hypertension*, I write to you from the editorial office of PLOS ONE to inform you of concerns related to duplicated content in articles published by your journals. We have been following up on concerns of overlapping text and duplicate Western blots within the following PLOS ONE article: [1] Berberine Improves Kidney Function in Diabetic Mice via AMPK Activation https: doi.org 10.1371 journal.pone.0113398 Received: June 9, 2014; Accepted: October 23, 2014; Published: November 19, 2014 It was initially brought to our attention that there is duplication of Western blot images between the PLOS ONE article and the following published papers: - [2] Brain Injury (Received 28 Oct 2013, Accepted 4 Jan 2015, Published online 20 Mar 2015) doi: 10.3109 02699052.2015.1004746: Figure 6b GAPDH is similar to Figure 2A AMPK in [1] - [3] Exp Mol Pathol (Received 24 Feb 2014, Accepted 10 Sep 2014, Available online 16 Sep 2014) doi:10.1016 j.yexmp.2014.09.006: Figure 5B GAPDH is similar to Figure 2A AMPK in [1]; Figure 5C Occludin is similar to Figure 2A LKB1 in [1] - [4] Korean J Physiol Pharmacol, (Received 7 Nov 2013, Accepted 3 Jan 2016) doi: 10.4196 kjpp.2016.20.4.325 RETRACTED: Figure 6B GAPDH is similar to Figure 2A AMPK [1] Please note that the KJPP paper has been retracted as a result of the content duplication issues. During the course of our follow up, we have discovered additional instances of possible duplication as follows: - [5] Gastroenterology and Hepatology (Accepted 6 Aug 2014, Accepted ms online 28 Aug 2014, Published 23 Feb 2015) doi:10.1111 jgh.12723: Figure 1d AMPK is similar to Figure 2A AMPK in [1], Figure 3a iNOS is similar to Figure 2A LKB1 in [1] - [6] Acta Pharmacologica Sinica (Received 4 Mar 2014, Accepted 28 July 2014, Published 17 Nov 2014) doi: 10.1038 aps.2014.88: Figure 1 A and B bar charts are similar to Figure 1 A and B bar charts in [1], Figure 1E AMPK is similar to Figure 2A AMPK in [1], Figure 1E p-AMPK is similar to Figure 2A P-AMPK in [1], Figure 1E bar chart is similar to the Figure 2A bar chart in [1] - [7] Asian Pac J Cancer Prev (Published Jan 2014) doi: 10.7314 APJCP.2014.15.2.677: Figure 3A GAPDH is similar to Figure 2A AMPK in [1] and p27 kip1 is similar to Figure 2A P-AMPK in [1] - [8] Clinical and Experimental Hypertension (Received 15 Sep 2015, Accepted 24 Nov 2015, Published 5 May 2016) doi: 10.3109 10641963.2015.1131288: Figure 2A LKB1 and P-AMPK are similar to Figure 2A LKB1 and P-AMPK in [1], Figure 2B P-AMPK and AMPK are similar to Figure 2B P-AMPK and AMPK in [1], Figure 2A and B bar charts appear similar in both articles. Figures 1A, B and C including Western blots and charts appear similar in both articles. Figure 6C AMPK is similar to Figure 2A AMPK in [1] (note that authors Min Hu and Bo Liu may be the same as authors on the retracted KJPP paper above) Articles [5], [6], [7], and [8] contain various amounts of duplicated text in the Results sections when compared to the PLOS ONE article. Note that there may be other instances of duplicated data and or text between the above articles aside from those affecting the PLOS ONE article. For at least some of the duplicated text, it appears that some of the manuscripts were under consideration at overlapping times. We have been informed that an external biotechnology company conducted the Western blot experiments and provided the raw blots to the authors for the PLOS ONE paper. How the duplications in text and figures arose remains unresolved. Although our correspondence with the authors about this matter is ongoing, we have determined that it is appropriate to issue a retraction of the PLOS ONE article, and the retraction notice will provide details of the similarities in content with the above-listed articles. We will also report this matter to the PLOS ONE authorsu2019 institution. I hope that the information provided above is helpful. If you have any questions in the course of any follow up on this matter, please do not hesitate to get in touch. Best wishes ,