Gene expression profiling of murine wounds revealed marked, sustained induction of cardiac ankyrin repeat protein (CARP, ankrdl), with diminished expression in diabetic mice. Overexpression of CARP in wounds by adenoviral gene transfer produces a remarkable increase in neovascularization and thereby enhances wound healing. Under the previous aims, we established these effects in several animal models and showed that CARP induction occurs in microvascular endothelial cells (MVEC), in part, through TGF-IS mediated activation of a p38 kinase pathway. CARP also protects endothelial cells from doxorubicin-mediated apoptosis, and it enhances MVEC migration, not proliferation. To avoid overlap, the proposed studies will concentrate on gain-of-function aspects of CARP. The proposed studies will extend the original aims by delineating the CARP activation pathway as well as identifying its binding partners and direct transcriptional targets. We will test the hypothesis that hyperglycemic, oxidative stress is related to CARP induction. Experiments will determine the influence of TNF-a and possible convergent signal mechanisms. Binding partners in vascular cells will be identified by yeast two- hybrid, protein microarray, and tandem affinity purification methods. EMSA and ChIP analysis will be used to identify the transcriptional activators in MVEC and smooth muscle cells (SMC). Expression profiling will be expanded to identify key targets in both MVEC and SMC. CARP has been suggested to be a mediator of mechanical stress in cardiac muscle. Experiments will test the significance of CARP'S putative nuclear and cytoplasmic interaction sites in SMC and MVEC by deletion and mutation of discrete, functional domains of the protein, read out as migratory and stress responses of targeted cells. Partners for these sites will be identified. The clinical relevance of CARP will be investigated in a series of rat healing studies in diabetic and ischemic wounds with a novel gene delivery system. These experiments will define the optimal dose for repair, and investigate the role of CARP in vessel regression and stabilization. If the benefits and mechanisms of this form of wound therapy are validated, they could lead to the development of CARP as a novel therapeutic and to the discovery of factors that mediate CARP-enhanced neovascularization.
| Zhong, Lin; Chiusa, Manuel; Cadar, Adrian G et al. (2015) Targeted inhibition of ANKRD1 disrupts sarcomeric ERK-GATA4 signal transduction and abrogates phenylephrine-induced cardiomyocyte hypertrophy. Cardiovasc Res 106:261-71 |
| Samaras, Susan E; Almodóvar-García, Karinna; Wu, Nanjun et al. (2015) Global deletion of Ankrd1 results in a wound-healing phenotype associated with dermal fibroblast dysfunction. Am J Pathol 185:96-109 |
| Baquerizo Nole, Katherine L; Yim, Elizabeth; Van Driessche, Freya et al. (2014) Wound research funding from alternative sources of federal funds in 2012. Wound Repair Regen 22:295-300 |
| Almodóvar-García, Karinna; Kwon, Minjae; Samaras, Susan E et al. (2014) ANKRD1 acts as a transcriptional repressor of MMP13 via the AP-1 site. Mol Cell Biol 34:1500-11 |
| Richmond, Nicholas A; Lamel, Sonia A; Davidson, Jeffrey M et al. (2013) US-National Institutes of Health-funded research for cutaneous wounds in 2012. Wound Repair Regen 21:789-92 |
| Chen, Billy; Zhong, Lin; Roush, Sarah F et al. (2012) Disruption of a GATA4/Ankrd1 signaling axis in cardiomyocytes leads to sarcomere disarray: implications for anthracycline cardiomyopathy. PLoS One 7:e35743 |
| Samaras, Susan E; Chen, Billy; Koch, Stephen R et al. (2012) 26S proteasome regulation of Ankrd1/CARP in adult rat ventricular myocytes and human microvascular endothelial cells. Biochem Biophys Res Commun 425:830-5 |
| Schultz, Gregory S; Davidson, Jeffrey M; Kirsner, Robert S et al. (2011) Dynamic reciprocity in the wound microenvironment. Wound Repair Regen 19:134-48 |
| Davidson, Jeffrey M (2010) Can scarring be turned off? Am J Pathol 176:1588-91 |
| Eming, Sabine A; Krieg, Thomas; Davidson, Jeffrey M (2007) Gene therapy and wound healing. Clin Dermatol 25:79-92 |
