LncRNA-NEAT1 from the competing endogenous RNA network promotes cardioprotective efficacy of mesenchymal stem cell-derived exosomes induced by macrophage migration inhibitory factor via the miR-142-3p/FOXO1 signaling pathway
Authors: Hanbin Chen
Meng Hou
Wenzheng Xia
Journal: Stem Cell Research & Therapy (2020)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: Cardiomyocytes apoptosis; Exosome; LncRNA-NEAT1/miR-142-3p/FOXO1 signaling pathway; MIF; Mesenchymal stem cells
Affiliations: Department of Radiation Oncology, First Affiliated Hospital, Wenzhou Medical University, No. 2 Fuxue Lane, Wenzhou, 325000, People's Republic of China., Department of Neurosurgery, Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine, Shanghai, China., Department of Radiation Oncology, First Affiliated Hospital, Wenzhou Medical University, No. 2 Fuxue Lane, Wenzhou, 325000, People's Republic of China. .

Abstract:  Aims: Extracellular vesicles, especially exosomes, have emerged as key mediators of intercellular communication with the potential to improve cardiac function as part of cell-based therapies. We previously demonstrated that the cardioprotective factor, macrophage migration inhibitory factor (MIF), had an optimizing effect on mesenchymal stem cells (MSCs). The aim of this study was to determine the protective function of exosomes derived from MIF-pretreated MSCs in cardiomyocytes and to explore the underlying mechanisms. Methods and results: Exosomes were isolated from control MSCs (exosome) and MIF-pretreated MSCs (exosomeMIF), and delivered to cardiomyocytes subjected to H2O2 in vitro. Regulatory long non-coding RNAs (lncRNAs) activated by MIF pretreatment were explored using genomics approaches. ExosomeMIF protected cardiomyocytes from H2O2-induced apoptosis. Mechanistically, we identified lncRNA-NEAT1 as a mediator of exosomeMIF by regulating the expression of miR-142-3p and activating Forkhead class O1 (FOXO1). The cardioprotective effects of exosomeMIF were consistently abrogated by depletion of lncRNA-NEAT1, by overexpression of miR-142-3p, or by FOXO1 silencing. Furthermore, exosomeMIF inhibited H2O2-induced apoptosis through modulating oxidative stress. Conclusions: Exosomes obtained from MIF-pretreated MSCs have a protective effect on cardiomyocytes. The lncRNA-NEAT1 functions as an anti-apoptotic molecule via competitive endogenous RNA activity towards miR-142-3p. LncRNA-NEAT1/miR-142-3p/FOXO1 at least partially mediates the cardioprotective roles of exosomeMIF in protecting cardiomyocytes from apoptosis. 

Retracted: No

PubPeer comments:
 This article lists 1 forward and 1 reverse primer for detecting NEAT1 transcript in human cells. Specifically: LncRNA-NEAT1 **F: 5u2032-GTACGCGGGCAGACTAACAC-3u2032 **,** R: 5u2032-TGCGTCTAGACACCACAACC-3u2032** Primer-blast ![file] Same results at InSilico-PCR ![file] Same results at Seek Blastn ![file] Of interest, the same Reverse primer has been used in 3 other studies, according to Google Scholar: ![file] Of note, the 1st and 2nd of Google Scholar results, what normally is called reverse primer, is written as  **reward** primer. Could the authors explain all these discrepancies? , 
 #2 Agreed. ![file] , 
 For reasons that are surely inscrutable to rank and file outsiders, Springer-Nature has suddenly, out of the blue, chosen to puff some Year of the Metal Rat (2020) u201chigh impactu201d papers that only it has published and no one else. https: www.springernature.com gp researchers campaigns chinese-research One does begin to wonder, though, as to whether it might be a lucky escape for all those other u201chigh impactu201d scientific publishers? A circular RNA paper championed by said publisher has been deconstructed here https: pubpeer.com publications 0B53FF5CB2ABF5495DE07A724B3CF6 This paper interweaves lncRNA with stem cells and exosomes. Is that hot or what? Pushes all the buttons. No wonder it is so high impact! Yay! Hot though it is, Condy could not help but notice a FACS plot being reused between two figures. ![image] You know sometime you re bound to leave her But for now you re going to stay In the year of the rat , 
 #3 Thank you for the second opinion. We are not finished yet. One of the two problematic issues in Fig. 3F of this SPRINGER-NATURE u201chigh impactu201d paper is that two differently labelled FACS plots are almost, but not quite, completely identical ![image] And the other problem is not just for Fig. 3F: Why does the metadata at top right of all the FACS plots in this paper always need to blanked out with white rectangles? Enquiring figure sleuth minds want to know. , 
 #4 None of the FACS plots in this paper have any cells in their top right corners. In fact some do but all the plots have a white rectangle placed in the top corner. This is presumably hiding some sort of labelling? Here is one example. Note that a row of plotted points are truncated by the lower edge of the white rectangle as indicated by the red arrows, seen most clearly in the sharpened image to right. ![image] , 
 On 20th May 2020 Springer Nature signed up to the Declaration on Research Assessment (DORA). https: en.wikipedia.org wiki San_Francisco_Declaration_on_Research_Assessment And yet here they are in 2021 still using u201chigh impactu201d as a proxy for a paperu2019s quality. Hypocrites. Just stop it, stop it already. And here is another top quality high impact FACS reuse, again cunningly disguised with a few different spots ![image] But chicks from rotten eggs do not proceed, Nor is a hypocrite a saint indeed. The rotten egg, though underneath the hen, If crack d, stinks, and is loathsome unto men. Nor doth her warmth make what is rotten sound; What s rotten, rotten will at last be found. The hypocrite, sin has him in possession, He is a rotten egg under profession. John Bunyan , 
 Little evidence is shown to support the assertion that  exosomes were isolated specifically, and characterization is done only for one unspecified sample and one unspecified condition. Figure 1A shows two particles by TEM that are around 200 nm and do not display the typical dehydration artifact of vesicles.  Exosomes are also usually described as less than 150 nm. In Figure 1B, nanoparticle tracking is not specific for exosomes, as stated here, or even fro other extracellular vesicles, EVs. Figure 1C, single bands of Western blots in Figure 1C do not give context, and CD63 does not show the characteristic smear. None of the markers is specific for exosomes versus other EVs. No negative markers are shown. Like many other publications that claim to describe experiments with MSC exosomes, this article does not show the purity of exosomes, the absence of other EVs or the influence of co-isolated proteins. Cell health and even viability is not monitored, important if cells were really grown in absence of serum. , 
 Dear Condylocarpon Amazonicumuff1a Sorry for missing your previous comment. Thank you for pointing out this and your rigorous scientific attitude. This will be serving as a good and serious information for us in proper data presentation. We will investigate the issue thoroughly and will deal with it as soon as possible. Regards Meng Hou , 
 #8 Thank you for replying Dr. Hou. The issues are quite serious, especially when a FACS plot is reused with a few different spots for two different experiments. In addition, #7 brings up some additional points. I have checked the CD63 antibody ab59479 quoted in your paper and the displayed blot of the supplier is strongly smeared. I have never done a blot for this protein, but *Russula* suggests that smearing is to be expected. A second supplier also provides a smeared blot for CD63. Below, your Fig. 1C is compared to the images from these CD63 antibody suppliers. ![image] ,