1. The novel gene CDKL-1 potentially signals for the development of the cardiovascularture CDKL-1 ?

2. Meeting 22 nd April 2015 Agenda RECAP REPEATABLE? BIOINFORMATICS 1) Possible Pathway 2) Protein-Protein Interactions- new questions and avenues for research Summer Placements

3. CDKL-1 expression through development 72 hpf 24 hpf 48hpf Tricaine Fig. 3 72 hpf 24 hpf 48hpf Tricai ne Fi g. 3 24 hpf 48 hpf 72 hpf 24 hpfNotochord Hypochord 48 hpf

4. CDKL-1 expression in hypoxia model (vhl mutant) embryos. vhl + vhl - vhl +/+ (4/19)vhl +/- (5/19) vhl -/- (5/19) Homozygous vhl mutant expected phenotypic ratios of a monohybrid cross (the vhl gene exhibits codominance).

5. vhl -/- vhl +/+ vhl -/- transverse Notochord Dorsal aorta Posterior Cardinal vein Our hypothesis: CDKL1 is involved in signalling processes leading to blood vessel development

6. Potential pathway or mechanism Involvement of Shh/Notch Hsu et al., 2011 Hsu et al., 2011 demonstrated Shh reduced expression in CDKL-1 morpholino (panel H)

7. CDKL-1 shh VEGF NOTCH BLOOD VESSEL GROWTH CDKL-1 shh NOTCH -ve feedback VEGF Potential pathway or mechanism Shh inhibitor (cyclopamine) CDKL1elevation (endothelial cells) http://www.ncbi.nlm.nih.gov/geoprofiles/966776 46 http://www.ncbi.nlm.nih.gov/geoprofiles/966776 46 VEGF effect on CDKL1 elevation (endothelial cells) http://www.ncbi.nlm.nih.gov/geoprofiles/611100 62 http://www.ncbi.nlm.nih.gov/geoprofiles/611100 62

8. CDKL1 Protein-Protein interactions

9. Hsp90 Glucocorticoid receptor chaperone FKBP5 is one of Hsp90s GR co chaperones.

10. TRIP6 (Thyroid hormone receptor binding protein) Contains a zinc finger LIM domain. LIM domain divided into 14 classes TRIP6 is in the Zyxin class. Zyxin are non enzymatic proteins bound to focal adhesions allowing for a scaffold for protein protein interactions of a signal cascade initiation at the membrane. Zyxin may function as a messenger in the signal transduction pathway that mediates adhesion-stimulated changes in gene expression NATURE CELL BIOLOGY | LETTER The LIMD1 protein bridges an association between the prolyl hydroxylases and VHL to repress HIF-1 activity. (Fox et.al 2012) - In contrast to LIMD1 it has been reported that TRIP6 can stabilise HIF (Bridge 2013- thesis)

11. Title von Hippel-Lindau (VHL) tumor suppressor protein deficiency effect on pancreatic islet cells OrganismMus musculus Title VEGF-A effect on endothelial cell line: time course (HG- U133A) OrganismHomo sapiens

12. Profile GDS3800 / 1429664_at / Cdkl1 Title von Hippel-Lindau (VHL) tumor suppressor protein deficiency effect on pancreatic islet cells Organis m Mus musculus VHL mutant and CDKL1 expression

13. Is CDKL1 regulating HIF through activation of the LIM domain proteins? If it is should we be concerned about conclusions we now draw from the VHL mutants?

14. EDIL3 EDIL3 (DEL1) Epidermal growth factor-like repeats and discoidin I-like domains (EDIL3), Angiogenic-Extracellular matrix-Glycoprotein secreted by endothelial cells. promotes adhesion of endothelial cells to extracellular matrix through interaction with the alpha-v/beta- 3 integrin receptor. High expression in Hepato cellular carcinoma Transcriptomic and genomic analysis of human hepatocellular carcinomas and hepatoblastomas.Luo JH, Ren B, Keryanov S, Tseng GC, Rao UN, Monga SP, Strom S, Demetris AJ, Nalesnik M, Yu YP, Ranganathan S, Michalopoulos G Hepatology. 2006 Oct; 44(4):1012-24. Angiogenesis The embryonic angiogenic factor Del1 accelerates tumor growth by enhancing vascular formation.Aoka Y, Johnson FL, Penta K, Hirata Ki K, Hidai C, Schatzman R, Varner JA, Quertermous T Microvasc Res. 2002 Jul; 64(1):148-61. Vessel wall remodeling Neovascularization of ischemic tissues by gene delivery of the extracellular matrix protein Del-1. Zhong J, Eliceiri B, Stupack D, Penta K, Sakamoto G, Quertermous T, Coleman M, Boudreau N, Varner JA J Clin Invest. 2003 Jul; 112(1):30-41. Del-1 gene transfer induces cerebral angiogenesis in mice.Fan Y, Zhu W, Yang M, Zhu Y, Shen F, Hao Q, Young WL, Yang GY, Chen Y Brain Res. 2008 Jul 11; 1219():1-7. Exosome Secretion- releasing EDIL3 Bladder Cancer Exosomes Contain EDIL-3/Del1 and Facilitate Cancer Progression Carla J. Beckham a,,,Jayme Olsen b, Peng-Nien Yin a, Chia-Hao Wu a, Huei-Ju Ting d, Fred K. Hagen c,Emelian Scosyrev a, Edward M. Messing a, Yi-Fen The hurnal of Urology; Sep 18-22, 2013 ; Current thinking - Exosomes have the potential as Biomarkers. Ranked 73 rd out f 83 genes isolated in our study!!!!!!!!