OC 16.1 - Multi-ancestry GWAS for venous thromboembolism identifies novel loci after validation in zebrafish

Sunday, June 23, 2024

09:30 – 09:45 ICT

Room: 111 A-C

Co-Author(s)

Anna R. Dahlgren, PhD

Postdoctoral Research Fellow
University of Michigan
Ann Arbor, Michigan, United States
BW

Brooke Wolford

Postdoctoral Research Fellow
University of Michigan
Ann Arbor, Michigan, United States
YZ

Yakun Zhao

Research Lab Technician Associate
University of Michigan
Ann Arbor, Michigan, United States
IS

Ida Surakka

Research Area Specialist Lead
University of Michigan
Ann Arbor, Michigan, United States
KW

Kuan-Han Wu

PhD candidate
University of Michigan
Ann Arbor, Michigan, United States
XY

Xinge Yu

Research Investigator
University of Michigan
Ann Arbor, Michigan, United States
Catherine E. Richter, MS

Research Specialist
University of Michigan
Ann Arbor, Michigan, United States
CW

Cristen Willer

Associate Professor
University of Michigan
Ann Arbor, Michigan, United States

Presenting Author(s)

Jordan Shavit, MD, PhD

Professor
University of Michigan
Ann Arbor, Michigan, United States

Background: Venous thromboembolism (VTE) is a prominent cause of morbidity and mortality with over 900,000 individuals affected each year in the U.S. While a variety of factors contribute to VTE risk, 30-40% of that risk is heritable.

Aims: To identify genes associated with VTE risk and validate in an animal model.

Methods: We performed a multi-ancestry genome-wide association study (GWAS) by meta-analyzing results for VTE across nine international cohorts of the Global Biobank Meta-analysis Initiative (GBMI, 27,987 cases and 1,035,290 controls). Genes near significant loci were identified. The orthologs of six prioritized genes (F7, STAB2, RASIP1, TC2N, TSPAN15, and PLCG2) were knocked down in 3-day-old zebrafish embryos using CRISPR/Cas9. A CRISPR/Cas9 knockout of the TSPAN15 ortholog (tspan15b) and a plasmid overexpression model were developed. Time to occlusion (TTO) after laser-mediated endothelial injury was evaluated.

Results: We identified 38 genome-wide significant loci. Of the six prioritized genes, through knockdown and effects on TTO in embryos, we found significant evidence for a role of RASIP1 and TC2N in VTE, and suggestive evidence for PLCG2 and TSPAN15. Individual knockdowns of the two potential TSPAN15 orthologs (tspan15a and tspan15b) revealed that depletion of tspan15b affected TTO in embryos, indicating it as the ortholog. Overexpression of the tspan15b plasmid in wild-type larvae caused a significant increase in TTO (Median(control)=11sec; Median(plasmid)=13sec;P < 0.05), while overexpression in protein C mutants caused an even greater increase in TTO (Median(control)=35.5sec; Median(plasmid)=57sec;P < 0.01).

Conclusion(s): We found evidence for three new genes which have not been previously identified or validated as VTE modifiers. TSPAN15 might not only play a role on its own, but also have additive effects in the context of protein C deficiency. These data demonstrate the power of zebrafish to quickly validate and study GWAS loci. Further characterization may provide insight into the variable penetrance of thrombosis and novel insight into coagulation cascade regulation.