This project will focus on the development and in vivo characterization of small molecule inhibitors of HOXA13 as a therapy for reducing tumor vascularization. During embryogenesis, HOX proteins direct tissue and organ development by regulating clusters of genes whose products control tissue specification and differentiation. We have established that HOXA13 regulates a cluster of pro-vascular genes necessary for vascular development in the developing embryo. More importantly, we have determined that many of the same provascular genes are regulated by HOXA13 in the developing vasculature of hepatocarcinomas. Because HOXA13 functions in a pleiotropic manner to regulate vascular development, I hypothesize that the disruption of HOXA13's transcriptional function would effectively inhibit tumor vascularization by simultaneously affecting the expression of a cohort genes required for tumor vasculogenesis. Recognizing that DNA binding is required for HOXA13's transcriptional function, we developed a small molecule screen to identify compounds capable of preventing HOXA13 from interacting with its target DNA sequence. From this screen, we identified Compound B (CpdB), a small molecule that can partially inhibit HOXA13's ability to regulate a group of provascular genes. Through careful modifications to CpdB's initial structure, I hypothesize that the efficacy of this small molecule can be improved to inhibit HOXA13 function in developing tumors. To test this hypothesis, the following specific aims will be accomplished:
Aim 1. Determine the specificity of CpdB for HOXA13.
Aim 2. Quantitate the affinity of CpdB for HOXA13 and determine which amino acid residues within the HOXA13 DNA binding domain are contacted by CpdB.
Aim 3. Determination of structure activity relationships for HOXA13 and CpdB, optimization of CpdB potency, and improvement of CpdB solubility.
Aim 4. Determine the efficacy of CpdB analogs to perturb HOXA13-regulated provascular gene expression in developing tumors.

Public Health Relevance

The proposed investigations will develop a newly identified series of HOXA13 small molecule antagonists for their potential to inhibit vasculogenesis. Applications of these findings will translate into new therapies to prevent tumor vascularization and growth.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
1R01CA131458-01A1
Application #
7578163
Study Section
Drug Discovery and Molecular Pharmacology Study Section (DMP)
Program Officer
Lees, Robert G
Project Start
2009-01-01
Project End
2013-11-30
Budget Start
2009-01-01
Budget End
2009-11-30
Support Year
1
Fiscal Year
2009
Total Cost
$329,510
Indirect Cost
Name
Oregon Health and Science University
Department
Genetics
Type
Schools of Medicine
DUNS #
096997515
City
Portland
State
OR
Country
United States
Zip Code
97239
Carlson, Hanqian L; Quinn, Jeffrey J; Yang, Yul W et al. (2015) LncRNA-HIT Functions as an Epigenetic Regulator of Chondrogenesis through Its Recruitment of p100/CBP Complexes. PLoS Genet 11:e1005680
Turner, Matthew; Zhang, Yonghong; Carlson, Hanqian L et al. (2015) Chemical shift assignments of mouse HOXD13 DNA binding domain bound to duplex DNA. Biomol NMR Assign 9:267-70
Pattawong, Ommidala; Tan, Darlene Q; Fettinger, James C et al. (2013) Stereocontrol in asymmetric ?-lactam syntheses from imines and cyanosuccinic anhydrides. Org Lett 15:5130-3
Tan, Darlene Q; Younai, Ashkaan; Pattawong, Ommidala et al. (2013) Stereoselective synthesis of ýý-lactams from imines and cyanosuccinic anhydrides. Org Lett 15:5126-9
Tan, Darlene Q; Atherton, Amy L; Smith, Austin J et al. (2012) Synthesis of a ?-lactam library via formal cycloaddition of imines and substituted succinic anhydrides. ACS Comb Sci 14:218-23
Tan, Darlene Q; Martin, Kevin S; Fettinger, James C et al. (2011) Ammonia synthons for the multicomponent assembly of complex gamma-lactams. Proc Natl Acad Sci U S A 108:6781-6
Zhang, Yonghong; Larsen, Christine A; Stadler, H Scott et al. (2011) Structural basis for sequence specific DNA binding and protein dimerization of HOXA13. PLoS One 6:e23069
Perez, Wilma D; Weller, Crystal R; Shou, Siming et al. (2010) Survival of Hoxa13 homozygous mutants reveals a novel role in digit patterning and appendicular skeletal development. Dev Dyn 239:446-57
Zhang, Yonghong; Thornburg, Chelsea K; Stadler, H Scott et al. (2010) Backbone chemical shift assignments of mouse HOXA13 DNA binding domain bound to duplex DNA. Biomol NMR Assign 4:97-9
Zhang, Yonghong; Thornburg, Chelsea K; Stadler, H Scott et al. (2009) (1)H, (15)N, and (13)C chemical shift assignments of mouse HOXA13 DNA binding domain. Biomol NMR Assign 3:199-201