Alternative polyadenylation of single cells delineates cell types and serves as a prognostic marker in early stage breast cancer

PLoS One. 2019 May 17;14(5):e0217196. doi: 10.1371/journal.pone.0217196. eCollection 2019.

Abstract

Alternative polyadenylation (APA) in 3' untranslated regions (3' UTR) plays an important role in regulating transcript abundance, localization, and interaction with microRNAs. Length-variation of 3'UTRs by APA contributes to efficient proliferation of cancer cells. In this study, we investigated APA in single cancer cells and tumor microenvironment cells to understand the physiological implication of APA in different cell types. We analyzed APA patterns and the expression level of genes from the 515 single-cell RNA sequencing (scRNA-seq) dataset from 11 breast cancer patients. Although the overall 3'UTR length of individual genes was distributed equally in tumor and non-tumor cells, we found a differential pattern of polyadenylation in gene sets between tumor and non-tumor cells. In addition, we found a differential pattern of APA across tumor types using scRNA-seq data from 3 glioblastoma patients and 1 renal cell carcinoma patients. In detail, 1,176 gene sets and 53 genes showed the distinct pattern of 3'UTR shortening and over-expression as signatures for five cell types including B lymphocytes, T lymphocytes, myeloid cells, stromal cells, and breast cancer cells. Functional categories of gene sets for cellular proliferation demonstrated concordant regulation of APA and gene expression specific to cell types. The expression of APA genes in breast cancer was significantly correlated with the clinical outcome of earlier stage breast cancer patients. We identified cell type-specific APA in single cells, which allows the identification of cell types based on 3'UTR length variation in combination with gene expression. Specifically, an immune-specific APA signature in breast cancer could be utilized as a prognostic marker of early stage breast cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions / genetics*
  • Biomarkers, Tumor / analysis
  • Breast Neoplasms / diagnosis*
  • Breast Neoplasms / genetics
  • Cell Proliferation
  • Female
  • Gene Expression Profiling
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • Kidney Neoplasms / diagnosis*
  • Kidney Neoplasms / genetics
  • MicroRNAs / genetics*
  • Polyadenylation*
  • Prognosis
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • Sequence Analysis, RNA
  • Single-Cell Analysis / methods*
  • Tumor Microenvironment

Substances

  • 3' Untranslated Regions
  • Biomarkers, Tumor
  • MicroRNAs
  • RNA, Messenger

Grants and funding

This study was supported by grants from the National Research Foundation of Korea (NRF-2017R1A6A3A11027978 to N.K., NRF-2017M3C9A6044636 and NRF-2016R1A5A1011974 to H.L. and NRF-2017M3A9A7050803 to W.P.) and from the Ministry of Food and Drug Safety of Korea (16173MFDS004 to W.P.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. W.P. is employed by a commercial company, GENINUS Inc, who provided support in the form of salary for W.P., but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific role of this author is articulated in the ‘author contributions’ section.