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Direct Conversion of Oral Epithelial Cells to Progenitor Dental Mesenchymal Cells by a Novel microRNA‐Transcription Factor Pathway
Author(s) -
Mansaray Maurisa N,
Sharp Thad,
Li Xiao,
Huojun Cao,
Amendt Brad
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.902.4
Subject(s) - mesenchyme , reprogramming , microbiology and biotechnology , biology , regeneration (biology) , microrna , progenitor cell , mesenchymal stem cell , stem cell , transcription factor , dental pulp stem cells , cell , genetics , gene
There are many protocols used for regeneration therapies to regenerate tissues and structures. To date, tooth regeneration practices rely heavily on the use of stem cells, however, these can be difficult to obtain and culture. We have previously introduced a new method of cellular reprogramming by over expression of the transcription factor, Pitx2 and microRNA‐200a‐3p . We now demonstrate that over expression of the Paired Box 9 ( Pax9 ) gene and inhibition of miR‐200a using the Plasmid‐based miRNA Inhibitor System ( PMIS‐miR‐200a ), converts oral epithelial cells to progenitor dental mesenchyme cells. Transgenic DNA was introduced using the lentivirus packaging system. The microRNA inhibitor system PMIS‐miR‐200a was constructed by NaturemiRI. A two‐staged approach is used to generate stable oral epithelial cells expressing PAX9 , cells were sorted and after several weeks were transduced with PMIS‐miR‐200a , to inhibit miR‐200a activity. miR‐200a regulates Wnt and TGFß signaling and inhibition of miR‐200a allows for cells to convert to a progeny phenotype in the presence of PAX9 . These cells express early dental mesenchyme and stem cell markers making them suitable for tooth regeneration therapies. Current experiments are using reprogrammed dental epithelial and mesenchyme cells to form a tooth organ. These data demonstrate a new method for cell reprogramming and tissue regeneration. Support or Funding Information Support for this research was provided from grant DE013941 and DE018885 from the National Institute of Dental and Craniofacial Research.