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2 results on '"Kitani, Tomoya"'

1. Single-Cell RNA Sequencing of Human Embryonic Stem Cell Differentiation Delineates Adverse Effects of Nicotine on Embryonic Development.

Author

Guo, Hongchao, Tian, Lei, Zhang, Joe Z, Kitani, Tomoya, Paik, David T, Lee, Won Hee, and Wu, Joseph C

Subjects
Myocytes, Cardiac, Humans, Calcium, Reactive Oxygen Species, Nicotine, Gene Expression Profiling, Sequence Analysis, RNA, Cell Communication, Cell Cycle, Cell Differentiation, Gene Expression Regulation, Developmental, Embryonic Development, Single-Cell Analysis, Transcriptome, Human Embryonic Stem Cells, differentiation, embryonic development, embryonic stem cells, nicotine, single-cell RNA sequencing, smoking, Gene Expression Regulation, Developmental, Myocytes, Cardiac, Sequence Analysis, RNA, Biochemistry and Cell Biology, Clinical Sciences
Abstract

Nicotine, the main chemical constituent of tobacco, is highly detrimental to the developing fetus by increasing the risk of gestational complications and organ disorders. The effects of nicotine on human embryonic development and related mechanisms, however, remain poorly understood. Here, we performed single-cell RNA sequencing (scRNA-seq) of human embryonic stem cell (hESC)-derived embryoid body (EB) in the presence or absence of nicotine. Nicotine-induced lineage-specific responses and dysregulated cell-to-cell communication in EBs, shedding light on the adverse effects of nicotine on human embryonic development. In addition, nicotine reduced cell viability, increased reactive oxygen species (ROS), and altered cell cycling in EBs. Abnormal Ca2+ signaling was found in muscle cells upon nicotine exposure, as verified in hESC-derived cardiomyocytes. Consequently, our scRNA-seq data suggest direct adverse effects of nicotine on hESC differentiation at the single-cell level and offer a new method for evaluating drug and environmental toxicity on human embryonic development in utero.

Published
2019

2. The DEAD-box RNA-binding protein DDX6 regulates parental RNA decay for cellular reprogramming to pluripotency

Author

Kami, Daisuke, Kitani, Tomoya, Nakamura, Akihiro, Wakui, Naoki, Mizutani, Rena, Ohue, Masahito, Kametani, Fuyuki, Akimitsu, Nobuyoshi, and Gojo, Satoshi

Subjects
0301 basic medicine, Cytoplasm, RNA, Untranslated, RNA Stability, lcsh:Medicine, Gene Expression, RNA-binding protein, Biochemistry, DEAD-box RNA Helicases, Animal Cells, Gene expression, Medicine and Health Sciences, Small interfering RNAs, RNA, Small Interfering, lcsh:Science, Connective Tissue Cells, Multidisciplinary, Chemistry, Stem Cells, Gene Expression Regulation, Developmental, Non-coding RNA, Cellular Reprogramming, RNA Helicase A, Cell biology, Precipitation Techniques, Nucleic acids, Ribonucleoproteins, Connective Tissue, Cellular Types, Anatomy, Cellular Structures and Organelles, Reprogramming, Research Article, Epithelial-Mesenchymal Transition, Induced Pluripotent Stem Cells, Research and Analysis Methods, 03 medical and health sciences, Proto-Oncogene Proteins, microRNA, Genetics, Immunoprecipitation, Humans, Messenger RNA, Natural antisense transcripts, lcsh:R, RNA, Biology and Life Sciences, Cell Biology, Fibroblasts, Gene regulation, MicroRNAs, 030104 developmental biology, Biological Tissue, lcsh:Q
Abstract

Cellular transitions and differentiation processes require mRNAs supporting the new phenotype but also the clearance of existing mRNAs for the parental phenotype. Cellular reprogramming from fibroblasts to induced pluripotent stem cells (iPSCs) occurs at the early stage of mesenchymal epithelial transition (MET) and involves drastic morphological changes. We examined the molecular mechanism for MET, focusing on RNA metabolism. DDX6, an RNA helicase, was indispensable for iPSC formation, in addition to RO60 and RNY1, a non-coding RNA, which form complexes involved in intracellular nucleotide sensing. RO60/RNY1/DDX6 complexes formed prior to processing body formation, which is central to RNA metabolism. The abrogation of DDX6 expression inhibited iPSC generation, which was mediated by RNA decay targeting parental mRNAs supporting mesenchymal phenotypes, along with microRNAs, such as miR-302b-3p. These results show that parental mRNA clearance is a prerequisite for cellular reprogramming and that DDX6 plays a central role in this process.

Published
2018

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