miR‐4286 functions in osteogenesis and angiogenesis via targeting histone deacetylase 3 and alleviates alcohol‐induced bone loss in mice
Authors: Yixuan Chen
Kaiyang Wang
Pei Liu
Daoyu Zhu
Zisheng Ai
Youshui Gao
Junjie Gao
Junhui Yin
Yigang Huang
Pengbo Luo
Hongping Yu
Qianhao Yang
Journal: Cell Proliferation (2021)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: HDAC3; alcohol; angiogenesis; miR-4286; osteogenesis.
Affiliations: Department of Orthopedic Surgery Shanghai Jiao Tong University Affiliated Sixth People’s Hospital Shanghai China, The First Affiliated Hospital of Xiamen University Xiamen China, Institute of Microsurgery on Extremities Shanghai Jiao Tong University Affiliated Sixth People's Hospital Shanghai China, Department of Medical Statistics Tongji University School of Medicine Shanghai China

Abstract:  Objectives: Alcohol consumption is one of the leading factors contributing to premature osteopenia. MicroRNA (miRNA) coordinates a cascade of anabolic and catabolic processes in bone homeostasis and dynamic vascularization. The aim was to investigate the protective role of miR-4286 in alcohol-induced bone loss and its mechanism. Materials and methods: The effect of miR-4286 and alcohol on bone mesenchymal stem cells (BMSCs) and human umbilical vein endothelial cells (HUVECs) was explored via multiple in vitro assays, including cell proliferation, QPCR, Western blot, osteogenesis, angiogenesis etc miR-4286 directly regulated HDAC3 was investigated by luciferase reporter assay, and the function of HDAC3 was also explored in vitro. Moreover, alcohol-induced bone loss in mice was established to reveal the preventive effect of miR-4286 by radiographical and histopathological assays. Results: In vitro, ethanol dramatically inhibited the proliferation and osteogenesis of BMSCs, and substantially impaired the proliferation and vasculogenesis of HUVECs. However, a forced overexpression of miR-4286 within BMSCs and HUVECs could largely abolish inhibitory effects by alcohol. Furthermore, alcohol-induced inhibition on osteogenic and vasculogenic functions was mediated by histone deacetylase 3 (HDAC3), and dual-luciferase reporter assay showed that HDAC3 was the direct binding target of miR-4286. In vivo, micro-CT scanning and histology assessment revealed that miR-4286 could prevent alcohol-induced bone loss. Conclusions: We firstly demonstrated that miR-4286 might function via intimate osteogenesis-angiogenesis pathway to alleviate alcohol-induced osteopenia via targeting HDAC3. 

Retracted: No

PubPeer comments:
 This paper has a June 2007 Corrigendum, which can be found at: https: www.nature.com articles nature05812 In summary the following errors were found and corrected: * In Figure 1b, the IgG isotype control tracings for several of the plots (n=5) differ by 1 log in fluorescence intensity; a new version of Figure 1b are provided but for a different set of MAPC cells * in the original Fig. 1b, the plot for MHC I was a duplication of the FACS plot for Mac-1 * In the legend to Fig. 1b, the superscripts u2018ku2019 in MHC II (I-Ak) and MHC I (H- 2Kk) should be u2018bu2019. * In the Methods under u2018Differentiation culture and analysisu2019, the concentration of 109 M dexamethasone should be 0.05 mM. * In the Supplementary Information, the sequence of the primers rex-1 and oct-4 were incorrect; correct sequences are provided. However, what the Correction does not address is that six of the panels in Figures 1b and S1b from this Nature paper are also shown in a second paper by the same group, i.e. Yuehua Jiang et al. Experimental Hematology 30 (2002) 896u2013904, DOI: 10.1016 s0301-472x(02)00869-x. See: https: pubpeer.com publications 9F0F20DFB792B0606731DB6E20C3CE Although this Nature paper was published before the Exp Hem paper, the Exp Hem manuscript was submitted and accepted earlier than the Nature paper. The six panels duplicated between the two papers represent different MAPC cell lines, as can be learned from the Erratum belonging to the Exp Hematology paper, found at: https: www.exphem.org article S0301-472X(06)00305-5 abstract. This duplication came to light after a New Scientist investigation, as reported here: https: www.newscientist.com article dn14922-briefing-anatomy-of-a-stem-cell-controversy Since this duplication was not addressed in the Nature corrigendum, it might be of interest to show the figures next to each other and highlight the duplications. Note that the two sets of panels show in light blue colors appear to represent the same experiment, so their duplication is appropriate. ![file] , 
 Here is an additional finding for Figure 6, also not part of the Corrigendum. Could the authors please check? * Green boxes: a possible area of overlap between Figure 6 s panel a (control mouse, b-gal negative) and panel e (chimeric mouse, b-gal positive). ![file] , 
 Figure 6M: the top right of this panel appears to show some repetitive elements, highlighted here with yellow and orange rounded boxes. ![file] , 
 I am wiling to state that repetitive elements in various sections are a random coincidence. There must be a statistician who can support this. There is no reason for someone to manipulate one portion of a tissue section which does not have a bearing on the immuno-label in question. Unless I am very naive. , 
 #4 Can you find sections with similar repetitive elements? , 
 #4 I could be a label introduced by the recording software (timing, sample name,...) that was removed by copying the affected areas. While not necessarily fraud, it certainly is bad practice to do this without justification. , 
 #6 I suggest that publishing data without disclosure that it has been edited is de facto research misconduct, no matter the intent. A better solution to your hypothetical explanation (which was not given by the authors) would have been to leave the original label and explain why it was extraneous. This would allow readers to assess whether the data was actually what the experiments showed and not something else. However, the institution approaches investigations with an eye toward silencing critical discussions (aka u201cgratuitous reportsu201d) such as occur on PubPeer: u201dWe therefore want to protect our researchers against gratuitous reports or complaints via public forums even before an investigation could be carried out. u201d From: https: nieuws.kuleuven.be en content 2020 ku-leuven-completes-research-on-possible-breach-of-research-integrity ,