The lymphatic vasculature has been implicated in a variety of pathophysiological processes, including tumor metastasis, tissue inflammation, edema, impaired immune surveillance and disrupted fat absorption. Nevertheless, it is remarkable that modern medicine has yet to discover pharmacologically-tractable targets for the modulation of lymphatic vessel growth or function;largely because the molecular mechanisms that control lymphangiogenesis remain poorly understood. Our lab has shown that adrenomedullin (AM), a 52-amino acid peptide that is secreted by lymphatic endothelial cells (LECs), is essential for proper embryonic lymphatic vascular development and that fine-tuned control of AM dosage is critical for the regulation of both embryonic and pathological lymphangiogenesis. CXCR7 was originally identified as an AM receptor, but has more recently been studied for its functions as a decoy receptor for SDF-1. Our preliminary studies suggest that CXCR7 is expressed in developing LECs and that CXCR7-/- mice exhibit lymphatic phenotypes consistent with AM overexpression. Therefore, I propose to investigate the hypothesis that CXCR7 serves as a decoy receptor for AM which consequently provides fine-tuned control of AM-mediated proliferation of LECs. I will perform comprehensive phenotypic analysis of lymphangiogenesis in loss-of-function CXCR7-/- mice in order to define the role of CXCR7 during in vivo lymphangiogenesis. In addition, I propose to cross CXCR7-/- mice to mice that are haploinsufficient for AM in order test whether the decoy activity of CXCR7 is required for the control of AM- mediated lymphangiogenesis in vivo. I will also use both loss-of-function and gain-of-function in vitro models to elucidate whether CXCR7 dampens AM-mediated downstream signaling. LECs will be cultured and treated with AM for proliferation assays, immunoblot, and 3D spheroid sprouting assays to explore the role of CXCR7 on AM-mediated LEC proliferation, downstream signaling, and mature vessel formation. Results from this proposal will define the role of CXCR7 in AM signaling and lymphangiogenesis and may elucidate novel therapeutic targets for the treatment of poorly managed lymphatic vascular diseases.
Lymphedema, a chronic, progressive swelling of tissue is the most common of lymphatic diseases, and affects 140-250 million people worldwide, and yet, despite its prevalence, only rudimentary treatments are available;largely because the molecular mechanisms that control the development and function of the lymphatic system are not well understood. Studies proposed in this grant will clarify the mechanisms that regulate the development of this system and how the lymphatic vasculature contributes to disease progression. The basic knowledge gleaned from these studies may provide novel and unique approaches for the treatment of lymphatic vascular disease such as lymphedema.
|Klein, Klara R; Matson, Brooke C; Caron, Kathleen M (2016) The expanding repertoire of receptor activity modifying protein (RAMP) function. Crit Rev Biochem Mol Biol 51:65-71|
|Klein, Klara R; Caron, Kathleen M (2015) Adrenomedullin in lymphangiogenesis: from development to disease. Cell Mol Life Sci 72:3115-26|
|Wetzel-Strong, Sarah E; Li, Manyu; Klein, Klara R et al. (2014) Epicardial-derived adrenomedullin drives cardiac hyperplasia during embryogenesis. Dev Dyn 243:243-56|
|Klein, Klara R; Karpinich, Natalie O; Espenschied, Scott T et al. (2014) Decoy receptor CXCR7 modulates adrenomedullin-mediated cardiac and lymphatic vascular development. Dev Cell 30:528-40|