Transcriptome-wide comparison of the impact of Atoh1 and miR-183 family on pluripotent stem cells and multipotent otic progenitors. Mus musculus

Hair cells (HCs) are the mechanoreceptors responsible for hearing and balance in the inner ear. Multiple factors cause HC loss including aging, noise exposure and genetic predisposition. A barrier to hearing restoration after HC loss is the inability of mammalian auditory HCs to spontaneously regenerate. Multiple factors are shown to be crucial for HC development and represent good candidates for regenerative studies. Atoh1 is widely accepted to be necessary and contextually sufficient for driving HC fate. Furthermore, the miRNA-183 family is known to be expressed at the time of HC differentiation, and mutations in miR-96 (one member of the family) can cause deafness both in mouse and human. Our goal is to identify and compare the impact of Atoh1/miR-183 family alone and in combination on the transcriptome of two different developmental cell models; mouse embryonic stem cells (mESCs) and immortalized multipotent otic progenitors (iMOPs) Overall design: mESCs & iMOPs were transfected with plasmid vectors carrying RFP alone (pT), Atoh1+RFP (pAT), miR-183 family+RFP (p183T) or Atoh1+miR-183 family+RFP (p183AT) for 48 hours, then RFP-positive cells were sorted using fluorescence activated cell sorting. Gene expression profiling was assessed using Affymetrix Mouse Gene ST Arrays and trancriptome analysis comparing each tranfection condition to the control pT condition was conducted. Three distinct biological samples per condition were included. Unmanipulated (untransfected control; UT) was also included.