RNA modifications are ubiquitous in biology and present in all classes of cellular RNAs including eukaryotic messenger and long non-coding RNA. A large fraction of mammalian mRNA/lncRNA modifications are also known to be reversible, highly dynamic, and occur in cell type and cell state dependent manner. The dynamic RNA epitranscriptomes, those involving N6-methyladenosine (m6A) in particular, are known to regulate many cellular activities including mRNA splicing, export, cytoplasmic localization, stability, translation activity, microRNA processing, immune tolerance, and to impact cellular processes including proliferation, development, circadian rhythm, and embryonic stem cell differentiation. Consider m6A in mRNA/lncRNA as an example, dedicated writers, erasers, and readers exist in human cells to orchestrate an additional layer of complex post-transcriptional gene expression regulation. Emerging new functions of RNA modifications are expected to follow, with significant implications on many aspects of human health and disease. Despite high potentials and promises, current epitranscriptome studies are significantly hampered by the lack of technologies that enable quantitative mapping of any type of mRNA/lncRNA modifications at high resolution and high sensitivity. This proposal will develop new methods that target five abundant mRNA/lncRNA modifications, namely m6A, 5-methylcytosine (m5C), N1-methyladenosine (m1A), pseudo uridine (?), and 2'O- methyls (Nm) for high-throughput sequencing at single-base resolution and suitable for low input RNA isolated from just hundreds to thousands of cells. New bioinformatics tools will be developed in order to facilitate data analysis. The general approaches proposed can be broadly applied to sequence RNA modifications in other RNA species including more abundant ribosomal RNA, transfer RNA, snRNA, and snoRNA as well as miRNA and piRNA. We will apply the newly developed methods to obtain base-resolution maps of RNA modifications in order to associate with human diseases, and to proof-of-principle studies in neuro-biology. Our proposed research will establish high-throughput, high-resolution, and high-sensitivity methods for epitranscriptome research in all biological areas.

Public Health Relevance

Dynamic RNA modifications (epitranscriptomes) have emerged as a new mechanism of post-transcriptional gene expression regulation. The proposed research will develop high-throughput, high-resolution, and high- sensitivity technologies for functional investigations of epitranscriptomes.

Agency
National Institute of Health (NIH)
Institute
National Human Genome Research Institute (NHGRI)
Type
Research Project with Complex Structure (RM1)
Project #
5RM1HG008935-03
Application #
9524833
Study Section
National Human Genome Research Institute Initial Review Group (GNOM)
Program Officer
Smith, Michael
Project Start
2016-09-27
Project End
2021-06-30
Budget Start
2018-07-01
Budget End
2019-06-30
Support Year
3
Fiscal Year
2018
Total Cost
Indirect Cost
Name
University of Chicago
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
005421136
City
Chicago
State
IL
Country
United States
Zip Code
60637
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Huang, Huilin; Weng, Hengyou; Sun, Wenju et al. (2018) Recognition of RNA N6-methyladenosine by IGF2BP proteins enhances mRNA stability and translation. Nat Cell Biol 20:285-295
Weng, Hengyou; Huang, Huilin; Wu, Huizhe et al. (2018) METTL14 Inhibits Hematopoietic Stem/Progenitor Differentiation and Promotes Leukemogenesis via mRNA m6A Modification. Cell Stem Cell 22:191-205.e9
Hsu, Phillip J; Fei, Qili; Dai, Qing et al. (2018) Single base resolution mapping of 2'-O-methylation sites in human mRNA and in 3' terminal ends of small RNAs. Methods :
Yao, Bing; Li, Yujing; Wang, Zhiqin et al. (2018) Active N6-Methyladenine Demethylation by DMAD Regulates Gene Expression by Coordinating with Polycomb Protein in Neurons. Mol Cell 71:848-857.e6
Li, Miaomiao; Zhao, Xu; Wang, Wei et al. (2018) Ythdf2-mediated m6A mRNA clearance modulates neural development in mice. Genome Biol 19:69
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Liu, Jun; Eckert, Mark A; Harada, Bryan T et al. (2018) m6A mRNA methylation regulates AKT activity to promote the proliferation and tumorigenicity of endometrial cancer. Nat Cell Biol 20:1074-1083
Shi, Hailing; Zhang, Xuliang; Weng, Yi-Lan et al. (2018) m6A facilitates hippocampus-dependent learning and memory through YTHDF1. Nature 563:249-253

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