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. 2018 Jul;138(7):1662-1665.
doi: 10.1016/j.jid.2018.02.016. Epub 2018 Feb 23.

A Human Stem Cell-Based System to Study the Role of TP63 Mutations in Ectodermal Dysplasias

Jason D Dinella  1 , Jiangli Chen  1 , Saiphone Webb  1 , Elaine Siegfried  2 , Alanna F Bree  3 , Senthilnath Lakshmanachetty  1 , Velmurugan Balaiya  1 , Maranke I Koster  4 , Peter J Koch  5
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A Human Stem Cell-Based System to Study the Role of TP63 Mutations in Ectodermal Dysplasias

Jason D Dinella et al. J Invest Dermatol. 2018 Jul.
No abstract available

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Conflict of interest statement

Conflict of Interest: The authors state no conflicts of interest

Figures

Figure 1
Figure 1. Generating iPSC and iPSC-derived keratinocytes from AEC patient skin
(a) Patient affected by AEC exhibiting severe skin erosions. The patient and his parents consented to the use of this image in this publication. (b) TP63 mutations in AEC patients occur mainly in exons 13-14, encoding putative protein-protein interaction domains. Approximate location of mutations in the AEC patient cells (F5 and E3) used in this manuscript are indicated by stars. The protein schematic shown is of ∆Np63α, the predominantly expressed TP63 isoform in human keratinocytes. DBD: DNA binding domain, OD: oligomerization domain, SAM: sterile alpha motif, PS: post-SAM domain. Phase contrast images of (c) human iPSC colony and (d) iPSC-derived keratinocytes. (e) Disease pathways associated with the TP63 mutations in E3 and F5 iPSC-K as determined by a Human Phenotype Ontology analysis of our RNAseq data (FDR; false discovery rate).
Figure 2
Figure 2. Characterization of desmosomal protein expression in AEC iPSC-K and AEC patient skin
Immunofluorescence analysis of a conisogenic pair of AEC and gene-corrected iPSC-K (F5, F5GC) in low calcium (a) and high calcium media for 24 hours (b-d). (a) TP63 and KRT14, (b) DSC3 and KRT14, (c) DSG3 and KRT14, and (d) DSG1/2 and KRT14 expression and localization. (e) Dispase assay showing accelerated fragmentation of AEC iPSC-K sheets in response to mechanical stress. (f) Treatment with the p38MAPK inhibitor SB202190 prevents AEC iPSC-K sheet fragmentation. (g) Western blot analysis for the indicated proteins of F5 and F5GC iPSC-K after exposure to high calcium conditions for 24 hours. Similar data for E3 and E3GC are shown in Supplemental Figure 2. Note downregulation of several proteins in AEC iPSC-K (JUP, 0.24; DSG3, 0.18; DSG1, 0.41; KRT1, 0.063; DSC3, 0.061; DSC1, 0.49). The ratio of pErk/Erk was increased 4.2 fold in AEC iPSC-K. (h-j) Immunofluorescence analysis of normal human skin and (k-m) AEC skin for (h,k) DSC3, (i, l) DSG1, and (j, m) DSC1. Arrowheads point towards normal expression while arrows point towards abnormal expression in panels h-j. Size bars: 10μm (a-d), 100μm (h-m)
See this image and copyright information in PMC

References

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