| 研究実績の概要 |
We studied the functions and potential roles of CD44 proteins in human skin. We for the first time identified up to 18 transcripts of CD44 in human skin epidermal keratinocytes and studied their roles in normal and disease skin development. To gain insight into CD44 function, we performed immunostaining of normal skin and various skin diseases. We found that CD44 variant expression tend to be reduced in various skin cancers, particularly squamous cell carcinoma but not in inflammatory diseases such as psoriasis and atopy dermatitis.
Using a keratinocytes cell line, we have been able to completely knock-out CD44 by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system through homology directed repair (HDR) and the cells expressed no CD44. To gain insight into pathways controlled by CD44, we performed Micro-array analysis using RNA from CD44 wild type and CD44 knockout cells. We found several genes that are controlled by CD44 proteins. Some important genes are either up- or down-regulated. Particularly, we found that knockdown of CD44 led to reduced expression of all 3 subunits of a certain laminin, which is important for maintaining structural integrity of human skin. We confirmed that the protein level is decreased in the cells.
Furthermore, we were able to establish keratinocyte cell lines expressing a specific CD44 molecule. These cells will be useful for identifying which CD44 protein have a particular function, because each CD44 molecule is thought to have a unique function.
Our study revealed important functions of CD44 in human skin.
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