Abstract
Background: Angiogenesis, forming new blood vessels from existing ones, is crucial in development, diabetes, and cancer. We found that Large Ribosomal subunit protein 17 (RpL17) played a key role in endothelial cell (EC) proliferation and angiogenesis. Reduced RpL17 (KD) promoted angiogenesis in human ECs and RpL17-deficient mouse retinas. RNA-sequencing and bioinformatics suggested a role for Early growth response 1(EGR1) transcription factor. Bioinformatics revealed Breast Cancer gene 1 (BRCA1) as a potential upstream regulator of RpL17, suggesting a novel pathway for angiogenesis. Hypothesis: BRCA1 interacts with RpL17 and upregulates EGR1 to stimulate vessel elongation in angiogenesis. Goal: Determine the role of BRCA1-RpL17 interaction in angiogenesis and their potential feedback loop via EGR1. Methods/Approach: RNA-sequencing of mouse ECs, whole retina mount staining (comparing wild-type and RpL17+/- mice). Tube formation assays with HUVECs to investigate angiogenesis after RpL17, BRCA1, and EGR1 knockdowns. Bioinformatics analysis with Python for RNA-seq data. Immunoblotting and RT-qPCR to measure protein and DNA expression. Immunofluorescence (IF) staining of HUVECs to observe BRCA1, RpL17, and EGR1 expression and localization. Preliminary Results: Whole retina mounts showed increased angiogenesis in RpL17-deficient mouse retinas compared to wildtype (p<0.009). HUVECs with RpL17 KD had increased junctions and longer branches than control HUVECs (p<0.0002). RNA-seq identified increased EGR1 expression (q<2.5x10 -8 ) in RpL17 KD ECs, confirmed with RT-qPCR (p<0.002). Bioinformatics predicted BRCA1 as a regulator of RpL17. HUVEC IF staining showed decreased RpL17 expression after BRCA1 KD. Angiogenesis assays of BRCA1 KD ECs exhibited an angiogenic phenotype like RpL17 KD, rescued following double BRCA1/RpL17 KD. Conclusion: BRCA1-RpL17 interaction stimulates angiogenesis and may be regulated by an EGR1 feedback loop, likely by EC proliferation. Detailed understanding of this pathway will provide new therapies to regulate angiogenesis.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have