Endothelial Notch signaling drives essential and diverse functions in angiogenesis. Abnormal Notch signaling is a significant contributor to many cardiovascular diseases; from blindness, stroke, and ischemic disease, to arteriovenous malformation (AVM) and tumor angiogenesis. Notch genes encode transmembrane receptors which undergo ligand-dependent cleavage of the intracellular domain; the intracellular domain then translocates to the nucleus, where it assembles transcriptional complexes to regulate the expression of downstream targets that carry out Notch function. Notch receptors respond to two classes of membrane bound ligands, Delta-like (Dll) and Jagged (Jag). Dll4 signals through Notch in normal and pathological angiogenesis to restrict sprout formation and promote perfusion, whereas Jag1 may have both Notch activating and inhibitory roles in endothelial cells. The transcriptional targets of Notch signaling drive many cellular functions that are critical for proper angiogenesis, yet our knowledge of Notch targets is currently limited. Defining these targets in the endothelium is needed to obtain a deeper knowledge of Notch angiogenic function and identifying the novel effectors will impact our ability to understand healthy vessels and human disease. Our preliminary studies highlight that Notch regulates several endothelial proteins that function in GPCR signaling, an unexplored area of endothelial Notch signaling. Our overall goal is to define endothelial genes regulated by Notch and use this knowledge to identify Notch effectors that regulate GPCR signaling and their mode of action in the vasculature. In this proposal, we will stimulate endothelial Dll4- or Jag1-Notch mediated transcription in vitro and identify Notch targets by RNAseq. A RiboTag mouse model will determine Notch-responsive endothelial genes in vivo. Combining data from the in vivo and in vitro analysis will lead to selection of candidate Notch targets that function in GPCR pathways and these will be validated as direct transcriptional targets. Using in vitro co- culture angiogenesis assays, we will determine if Notch-induced GPCR signaling molecules are required in order for Notch to regulate sprouting angiogenesis. One such candidate, Rnd1, a constitutively active GTPase, will be analyzed to test the hypothesis that Notch induces Rnd1 to regulate Rap-p120RasGAP- Ras signaling in endothelial cells. Rnd1, and other GPCR proteins induced by Notch, will be studied in mice to determine their roles in Notch regulated sprouting angiogenesis, hypoxia-induced retinopathy and arteriovenous malformation (AVM).

Public Health Relevance

The Notch signaling pathway is fundamental to proper vascular development and angiogenesis. We will study how Notch regulates angiogenesis and promotes proper formation and function of blood vessels. This will allow an understanding of how angiogenesis occurs in healthy individuals and human disease, and development of rationale strategies to repair retinal vascular disorders, inhibit tumors, control inflammatory conditions, and promote wound healing.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL112626-07
Application #
9616294
Study Section
Cardiovascular Differentiation and Development Study Section (CDD)
Program Officer
Gao, Yunling
Project Start
2012-07-17
Project End
2021-12-31
Budget Start
2019-01-01
Budget End
2019-12-31
Support Year
7
Fiscal Year
2019
Total Cost
Indirect Cost
Name
University of Illinois at Chicago
Department
Physiology
Type
Schools of Medicine
DUNS #
098987217
City
Chicago
State
IL
Country
United States
Zip Code
60612
Nowak-Sliwinska, Patrycja; Alitalo, Kari; Allen, Elizabeth et al. (2018) Consensus guidelines for the use and interpretation of angiogenesis assays. Angiogenesis 21:425-532
Dubeykovskaya, Zinaida A; Duddempudi, Phaneendra Kumar; Deng, Huan et al. (2018) Therapeutic potential of adenovirus-mediated TFF2-CTP-Flag peptide for treatment of colorectal cancer. Cancer Gene Ther :
Edwards, Andrew K; Glithero, Kyle; Grzesik, Peter et al. (2017) NOTCH3 regulates stem-to-mural cell differentiation in infantile hemangioma. JCI Insight 2:
Cuervo, Henar; Pereira, Brianna; Nadeem, Taliha et al. (2017) PDGFR?-P2A-CreERT2 mice: a genetic tool to target pericytes in angiogenesis. Angiogenesis 20:655-662
Tattersall, Ian W; Du, Jing; Cong, Zhuangzhuang et al. (2016) In vitro modeling of endothelial interaction with macrophages and pericytes demonstrates Notch signaling function in the vascular microenvironment. Angiogenesis 19:201-15
Munabi, Naikhoba C O; England, Ryan W; Edwards, Andrew K et al. (2016) Propranolol Targets Hemangioma Stem Cells via cAMP and Mitogen-Activated Protein Kinase Regulation. Stem Cells Transl Med 5:45-55
Westerterp, Marit; Tsuchiya, Kyoichiro; Tattersall, Ian W et al. (2016) Deficiency of ATP-Binding Cassette Transporters A1 and G1 in Endothelial Cells Accelerates Atherosclerosis in Mice. Arterioscler Thromb Vasc Biol 36:1328-37
Kangsamaksin, Thaned; Murtomaki, Aino; Kofler, Natalie M et al. (2015) NOTCH decoys that selectively block DLL/NOTCH or JAG/NOTCH disrupt angiogenesis by unique mechanisms to inhibit tumor growth. Cancer Discov 5:182-97
Verginelli, Federica; Adesso, Laura; Limon, Isabelle et al. (2015) Activation of an endothelial Notch1-Jagged1 circuit induces VCAM1 expression, an effect amplified by interleukin-1?. Oncotarget 6:43216-29
Kofler, Natalie M; Cuervo, Henar; Uh, Minji K et al. (2015) Combined deficiency of Notch1 and Notch3 causes pericyte dysfunction, models CADASIL, and results in arteriovenous malformations. Sci Rep 5:16449

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