A major molecular hallmark of cancer cells is the aberrant splicing of genes with antagonistic splice isoforms. One of such genes is the vascular endothelial growth factor-A (VEGF-A) that can be spliced as either canonical pro-angiogenic isoforms or anti-angiogenic isoforms. In most normal adult tissues, VEGF-A is predominantly spliced as the anti-angiogenic b isoforms, whereas the pro-angiogenic isoforms become dominant in cancer cells to promote the growth of new blood vessels. Since sustained angiogenesis is critical to tumor growth and metastasis, restoration of normal VEGF-A splicing of in tumors presents a new anti-tumor approach. Long-term objectives and specific aims: We propose to develop a novel anti-angiogenesis approach by manipulating VEGF-A splicing. Such method will rewire the natural splicing regulatory pathway that orchestrates angiogenesis, thus inhibiting tumor growth. There are two specific aims in this project: (1) Specifically modulate VEGF-A splicing to promote anti- angiogenic isoforms;(2) Determine the in vivo efficacy of VEGF-A splicing modulation as anti-angiogenic therapy. Research design and methods Using a new technique recently developed in our lab, we will generate novel engineered splicing factors (ESFs) to shift the splicing of VEGF-A from angiogenic isoforms to anti-angiogenic isoforms. The """"""""designer"""""""" ESFs will be expressed in metastatic breast and lung cancer cells, and the induction of anti-angiogenic VEGF- A splicing isoforms will be measured at mRNA and protein levels. As a complementary approach, we will use antisense oligonucleotides (ASOs) to shift the VEGF-A splicing toward anti-antigenic isoforms. We will further examine the response of the VEGF-A downstream signaling pathway to such splicing modulation. In addition, we will investigate whether the splicing modulation leads to the inhibition of angiogenesis using a variety of cell-based assays, and test the off-target effects of splicing modulation. We will further examine if the modulation of VEGF-A splicing can inhibit the tumor angiogenesis and growth in xenograft mouse model of human cancers. We will inject cancer cells pretreated with ESFs or ASOs into SCID mice to determine if the splicing modulation can inhibit the xenograft tumor formation and growth. Further we will directly treat the established xenograft tumors with intratumoral injection or systematic delivery of ESF or ASOs, and determine if such treatment can inhibit tumor progression. We will also combine splicing modulation of VEGF-A with other cancer therapies to see if they have synergistic effect to inhibit tumor progression in live animals.

Public Health Relevance

Cancer has become the leading cause of death worldwide. The growth and metastasis and cancers require sustained angiogenesis that is orchestrated by the balance of VEGF-A splicing isoforms. In this proposal, we will develop new techniques to modulate VEGF-A splicing, and thus to convert pro-angiogenic isoforms into anti-angiogenic isoforms in cancer cells. This work will provide the basis for novel anti-cancer therapies.

National Institute of Health (NIH)
National Cancer Institute (NCI)
Research Project (R01)
Project #
Application #
Study Section
Drug Discovery and Molecular Pharmacology Study Section (DMP)
Program Officer
Snyderwine, Elizabeth G
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of North Carolina Chapel Hill
Schools of Medicine
Chapel Hill
United States
Zip Code
Song, Xiaowei; Zeng, Zhenyu; Wei, Huanhuan et al. (2018) Alternative splicing in cancers: From aberrant regulation to new therapeutics. Semin Cell Dev Biol 75:13-22
Wei, Huan-Huan; Liu, Yuanlong; Wang, Yang et al. (2017) Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells. J Vis Exp :
Qi, Yangfan; Yu, Jing; Han, Wei et al. (2016) A splicing isoform of TEAD4 attenuates the Hippo-YAP signalling to inhibit tumour proliferation. Nat Commun 7:ncomms11840
Dominguez, Daniel; Tsai, Yi-Hsuan; Gomez, Nicholas et al. (2016) A high-resolution transcriptome map of cell cycle reveals novel connections between periodic genes and cancer. Cell Res 26:946-62
Dominguez, Daniel; Tsai, Yi-Hsuan; Weatheritt, Robert et al. (2016) An extensive program of periodic alternative splicing linked to cell cycle progression. Elife 5:
Wang, Yang; Wang, Zefeng (2016) Design of RNA-Binding Proteins: Manipulate Alternative Splicing in Human Cells with Artificial Splicing Factors. Methods Mol Biol 1421:227-41
Wei, Huanhuan; Wang, Zefeng (2015) Engineering RNA-binding proteins with diverse activities. Wiley Interdiscip Rev RNA 6:597-613
Szempruch, Anthony J; Choudhury, Rajarshi; Wang, Zefeng et al. (2015) In vivo analysis of trypanosome mitochondrial RNA function by artificial site-specific RNA endonuclease-mediated knockdown. RNA 21:1781-9
Tsai, Yihsuan S; Dominguez, Daniel; Gomez, Shawn M et al. (2015) Transcriptome-wide identification and study of cancer-specific splicing events across multiple tumors. Oncotarget 6:6825-39
Wang, Yang; Wang, Zefeng (2015) Efficient backsplicing produces translatable circular mRNAs. RNA 21:172-9

Showing the most recent 10 out of 20 publications