PKD Isoforms in Heart

描述（由申请人提供）：蛋白激酶D （PKD）是最近出现的一个具有重要心脏作用的酶家族。大多数研究都集中在PKD1（该酶家族的创始成员）磷酸化HDAC5， HDAC5是一种调节染色质重塑和肥大的II类组蛋白去乙酰化酶。PKD1也磷酸化CREB和肌聚蛋白（如心肌肌钙蛋白I和心肌肌球蛋白结合蛋白C），但它们在心脏生长反应中的作用尚未得到充分考虑。同样，许多PKD1底物也被PKD2或PKD3磷酸化，但PKD2或PKD3与PKD1合作调节心脏重塑的概念也未被考虑。我们的初步研究表明PKD1、2和3具有相似（但不完全相同）的体外底物

DESCRIPTION (provided by applicant): Protein kinase D (PKD) has recently emerged as a family of enzymes with important cardiac actions. Most studies have focused on PKD1 (the founding member of this enzyme family) which phosphorylates HDAC5, a class II histone deacetylase that regulates chromatin remodeling and hypertrophy. PKD1 also phosphorylates CREB and sarcomeric proteins (such as cardiac troponin I and cardiac myosin-binding protein C), but their role in cardiac growth responses has not adequately been considered. Similarly, many PKD1 substrates also are phosphorylated by PKD2 or PKD3, but the notion that PKD2 or PKD3 cooperate with PKD1 to regulate cardiac remodeling also has not been considered. Our preliminary studies showing that PKD1, 2, and 3 have similar (but not identical) in vitro substrate specificities, PKD1 and PKD2 are activated in a stimulus-specific manner in cardiomyocytes, and PKDs have both overlapping and non-redundant roles in mouse embryonic development and adult heart hypertrophy provide the rationale for the aims of this application. Aim #1 will use state-of-the- art peptide library screens to define PKD isoform substrate specificity. This aim also will use biochemical approaches and D Kinase Activity Reporter assays to identify activation mechanisms and signaling repertoires of individual PKD isoforms. We will use gene knockout and overexpression strategies (including with analog- sensitive PKD mutants) to identify PKD isoform-specific substrates and effectors in cardiomyocytes. Aim #2 will use PKD1, 2, and 3 single global knockout mice and PKD1/D2, PKD1/D3, and PKD2/D3 global and cell type specific double knockout mice to analyze the role of PKDs in cardiovascular development. Inducible adult cardiac specific PKD knockout mice and a transgenic analogue-sensitive PKD1 mutant mouse also will be generated to examine the role of PKDs in the adult myocardium. The overarching goal of these studies is to identify novel regulatory controls and cardiac actions of PKDs that can be targeted to prevent or slow the evolution of cardiac hypertrophy and heart failure phenotypes. PUBLIC HEALTH RELEVANCE: Protein kinase D (PKD) has recently emerged as a family of signaling enzymes with important cardiovascular actions, but efforts to consider PKD as a therapeutic target have been slowed by significant gaps in our understanding of the molecular controls of individual PKD family members and their distinct actions in the heart. Studies in this application will take advantage of innovative biochemical and molecular approaches to define the cardiac actions of PKD enzymes. These studies will be critical for the design and clinical application of PKD-targeted pharmaceuticals for the treatment of pathological cardiac remodeling.