Functional study of 14-3-3 protein epsilon (YWHAE) in keratinocytes: microarray integrating bioinformatics approaches

Abstract

Previously, we detected that 14-3-3 protein epsilon (YWHAE) was involved in the pathogenesis of atopic dermatitis (AD) and tyrosinase-mediated pigmentation. In this study, we aimed to identify critical factors associated with YWHAE in human keratinocytes using high-throughput screening (HTS) approaches to reveal its functions in skin. We overexpressed YWHAE in human HaCaT keratinocytes and then conducted serial HTS studies, including RNA sequencing integrated with antibody arrays and the implementation of bioinformatics algorithms. Cumulatively, these approaches identified several novel genes in keratinocytes associated with the function of YWHAE including KRT9, KRT1, KRT6C, BST2, CIB2, APH1B, ACTC1, IFI27, TUBA1A, CAPN6, UTY, MX2, and MAPK15, based on RNA sequencing data, and MAPK1, MMP2, TYK2, NOS3, and CASP3, based on antibody array data. In particular, CD37 is a unique gene that was detected and validated in all the methods applied in this study. By integrating the datasets obtained from these HTS studies and utilizing the strengths of each method, we obtained new insights into the functional role of YWHAE in skin keratinocytes. The approach used here could contribute to the clinical understanding of YWHAE-associated applications in the treatment of AD disease.

Abbreviations
DAVID	=	the database for annotation, visualization and integrated discovery
HTS	=	High-throughput screening
KEGG	=	Kyoto Encyclopedia of Genes and Genomes
PPI	=	protein-protein interactions

Communicated by Ramaswamy H. Sarma

Keywords:

• YWHAE
• microarray
• keratinocytes

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

This research was supported by a fund from the Public Project of Jiaxing City (No. 2018AY32042). Dr. Jun-Mo Yang was supported by a grant of the Korea Health Technology R&D Project through the Korean Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (grant number: HI17C0616) and by National R&D Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT and Future Planning (2017R1D1A1B03029114). Dr. Jae-Rin Lee was supported by Basic Science Research Program through NRF funded by the Ministry of Education (NRF-2018R1A6A3A11045540). Dr. Myong-Joon Hahn was supported by Basic Science Research Program through NRF funded by the Ministry of Education (2013R1A1A2013708) and Samsung Biomedical Research Institute grant (SMX1131691). Dr. Yong-Doo Park was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIP; Ministry of Science, ICT & Future Planning) (No. 2017R1A6A3A11033277).