1. NIH-R25 Summer Program for Undergraduate Research
Transcriptional Regulation of Heart Valve Development: The Roles of VEGF and NFAT Signaling
Ben Smood
University of Oregon
Stankunas Lab
NIH-R25 Summer Program for Undergraduate Research

2. Outline Significance Development
Known Roles of VEGF and NFAT signaling
Methods, Goals, and Plans
Questions

3. Significance Heart valve defects in 2-3% of population
Bicuspid aortic valve disease
Mitral valve prolapse
VSD
Single Set of Valves Effected
Egyptians: wisdom
VSD congenital-hypertension, heart failure, delayed growth and development
Starts beating at E18humans, pumping blood by E21
DSCR1 encodes inhibitor of calcineuron
DYRK1a promotes NFATc export

4. E9.5-10.5 EMT (major cushions seen at end)
E Cushion proliferation
elongation and remodeling
GET NEW FIGURE FROM BOOK

5. Valve Formation

6. VEGF and Valve Formation
Lambrechts and Carmeliet 2004

7. NFATc1: What’s Known
Zeini et al. 2009

8. NFAT Signaling Chang et al. 2004
Endocardial NFATc blocks VEGF in myocardium
Lose NFATc1 when mesenchymal cells; gone in EC after E13.5
NFATc1 for Emt only in OFT valves
Directly represses VEGF-A gene
No general angiogenic defects seen with inhibtion of VEGF signaling (VEGF-A is associated with angiogenesis though)
DSCR1 encodes inhibitor of calcineuron
DYRK1a promotes NFATc export
Chang et al. 2004

9. Slight differences have tremendous impacts
sFlt-sequesters VEGFR1 ligands (sensitive emt)
VEGFR2T-sequesters VEGFA, not VEGFB or plgf (not sensitive)
Placental growth factor

10. What are the transcriptional factors targeted by VEGF and NFATc1 signaling?

11. Aims: Understand the Roles of VEGF and NFATc1 in Development
Small molecule Inhibitors
Small molecule pharmagrade
Cyclosporine A, Cabozantinib, Sunitinib
TU-Tagging
Spatial & Temporal transcriptional events
Cabo inhibit vegfr2 phosphorylation
Suni VEGFR blocks

13. Double outlet right ventricle

14. A floxed-stop-UPRT line for modular cell-specific TU-tagging in mice
Uracil phosophoribosyltransferase
(ribose-5 phosphate to N1 nitrogen of Uracil. incorporated)

15. First method to covalently label RNA within specific cell types in the intact mouse

16. Acknowledgements Stankunas Lab SPUR Dr. Peter O’Day NIH-R25
Dr. Kryn Stanuknas
Dr. Scott Stewart
Vidusha Devasthali
Fern Bosada
Brynn Simeck
Alex Ackerberg
Maithri Sarangam
Alan Gomez
Cho Li
SPUR
Dr. Peter O’Day
NIH-R25
Peter O