Interfering with Protein-Protein Interaction for the Treatment of Leishmaniasis

干扰蛋白质-蛋白质相互作用治疗利什曼病

• 批准号: 7692203
• 负责人: DARIA MOCHLY-ROSEN
• 金额: $ 20.09万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-29 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7692203
• 关键词: Acute myocardial infarction; Affect; Animal Disease Models; Binding; Binding Proteins; Biological; Biological Assay; Biological Process; Cardiovascular Diseases; Cell Survival; Cell physiology; Cells; Cessation of life; Clinical Treatment; Cloning; Communicable Diseases; Complications of Diabetes Mellitus; Conserved Sequence; Consult; Country; Data; Death Rate; Development; Differentiation and Growth; Disease; Drug Binding Site; Drug Delivery Systems; Drug Industry; Drug resistance; Endemic Diseases; Enzymes; Esthesia; Event; Exploratory/Developmental Grant; Funding; Genome; Goals; Grant; Growth; Heart Transplantation; Homologous Gene; Human; Immune; In Vitro; Infection; Insulin; Isoenzymes; Knowledge; Laboratory Study; Lead; Leishmania; Leishmania leishmania; Leishmaniasis; Malignant Neoplasms; Methodology; Methods; Mus; Mutation; Myocardial Infarction; Nature; Pain; Parasites; Parasitology; Patients; Peptides; Pharmaceutical Preparations; Pharmacotherapy; Phase; Phase I Clinical Trials; Phase III Clinical Trials; Phosphotransferases; Plants; Protein Binding; Protein Biochemistry; Protein Kinase; Protein Kinase C; Proteins; Regulation; Reporting; Research; Resistance; Scaffolding Protein; Sequence Alignment; Sequence Homologs; Sequence Homology; Signal Pathway; Signal Transduction; Signaling Protein; Stroke; Structural Models; Techniques; Testing; Therapeutic; Therapeutic Agents; Time; Toxic effect; Yeasts; cell growth; chemotherapy; citrate carrier; computerized data processing; design; experience; in vitro activity; inhibitor/antagonist; interest; macrophage; neglect; novel; novel strategies; novel therapeutics; protein phosphatase inhibitor-2; protein protein interaction; receptors for activated C kinase; scaffold; tool; vector

DESCRIPTION (provided by applicant): Leishmaniasis is an endemic disease in tropical and subtropical regions of over 80 countries with more than 2 million new cases occurring annually http://www.who.int/tdr/diseases/leish/diseaseinfo.htm. Receptor for activated C kinase 1 (RACK1), a ubiquitous and highly conserved scaffold protein, binds to proteins involved in many signaling processes leading to cell survival, growth and differentiation. We have successfully generated short peptide regulators that either disrupt or enhance the interactions of RACK1 with these signaling proteins and thus regulate their biological effects. Little is known about the cellular function of the Leishmania homolog of RACK1, Leishmania homolog of receptors for activated C kinase, LACK. We believe that LACK, due to its highly conserved nature, is also a key scaffolding protein involved in essential signaling processes of the Leishmania parasite. We propose a novel approach, utilizing our well established methods of peptide design, to identify and characterize LACK- binding partners and to target protein-protein interactions between LACK and these Leishmaniasis- signaling enzymes. These peptides may serve as leads for the development of new therapeutics for the treatment of Leishmaniasis. Specifically, in this early feasibility proposal, we plan to: 1. Identify and characterize Leishmania signaling proteins that are LACK-binding partners (LACK-BP). 2. Rationally design peptides that selectively inhibit the interactions between the scaffold LACK and LACK-BP and test their activity towards Leishmania sp. proliferation and in vitro infection. To identify and characterize LACK-BP, we will utilize several methodologies including protein overlays (Far Western), in vitro pull-down assays, and expression interaction cloning. To design peptide modulators of protein-protein interactions, we will use sequences in RACK-BP that are required for RACK binding and identify homologous sequences in LACK-BP. Additionally, we will identify evolutionary conserved sequences in homologs of RACK-BP and use structural modeling to look for regions of homology for peptide design. Finally, we will identify short peptides derived from LACK that are likely to disrupt the binding and hence the signaling of LACK-BPs. The peptide modulators will be tethering to cell-permeable vectors and tested for their biological activity on proliferation of Leishmania promastigotes in culture and on Leishmania-infected murine macrophages. The peptides developed as part of the proposed project are likely to exert significant biological effects. Importantly, identification of LACK-binding proteins and peptides that interfere with their biological activities can help guide the development of new therapeutics for patients suffering from Leishmania infection. PROJECT NARRATIVE: Two million new cases of Leishmania infection are reported annually, with a resultant death rate of 3%. The current treatment for Leishmaniasis, chemotherapy, is highly toxic and ineffective. Our proposed studies will use a new approach to identify key players involved in Leishmania viability, allowing us to identify and create highly specific therapeutics that disrupt these cellular processes. This therapeutic approach is likely to provide a less toxic, highly specific and effective way of treating Leishmaniasis.

描述（由申请人提供）：利什曼病是80多个国家的热带和亚热带地区的一种流行疾病，每年有超过200万个新病例发生http://www.who.int/tdr/tdr/tdr/tdr/leish/leish/diseaseinfo.htm。活化C激酶1（RACK1）的受体是一种无处不在且高度保守的支架蛋白，与参与许多信号过程的蛋白质结合，导致细胞存活，生长和分化。我们成功地产生了短肽调节剂，以破坏或增强RACK1与这些信号蛋白的相互作用，从而调节其生物学作用。关于RACK1的利什曼原虫同源物的细胞功能，激活C激酶的受体同源物的细胞功能知之甚少。我们认为，由于其高度保守的性质，缺乏也是参与利什曼原虫寄生虫基本信号过程的关键脚手架蛋白。我们提出了一种新的方法，利用了我们确定的肽设计方法，以识别和表征缺乏结合伴侣，并靶向缺乏和这些利什曼病和这些利什曼病信号传导酶之间的蛋白质 - 蛋白质相互作用。这些肽可以作为开发利什曼病治疗新疗法的铅。具体而言，在这项早期的可行性建议中，我们计划：1。识别和表征缺乏结合伴侣的利什曼原虫信号传导蛋白（Laves-BP）。 2。理性设计的肽有选择地抑制脚手架缺乏和缺乏bp之间的相互作用，并测试其对利什曼原虫SP的活性。增殖和体外感染。为了识别和表征缺乏BP，我们将利用几种方法，包括蛋白质覆盖（远西方），体外下拉测定和表达相互作用克隆。为了设计蛋白质 - 蛋白质相互作用的肽调节剂，我们将使用RACK-BP中的序列，这些序列是RACK结合所需的，并识别缺乏BP中的同源序列。此外，我们将确定RACK-BP同源物中的进化保守序列，并使用结构建模来寻找用于肽设计的同源区域。最后，我们将确定由于缺乏可能破坏结合的短肽，因此缺乏BPS的信号传导。肽调节剂将束缚在细胞可渗透的载体上，并测试其在培养物和利什曼尼亚感染的鼠巨噬细胞中利什曼原虫的生物学活性。作为拟议项目的一部分而开发的肽可能会产生重大的生物学作用。重要的是，鉴定干扰其生物学活性的缺乏结合蛋白和肽可以帮助指导患有利什曼原虫感染的患者的新疗法。 项目叙述：每年有200万例利什曼原虫感染的新病例，导致死亡率为3％。当前对利什曼病的治疗方法是化学疗法，具有剧毒且无效。我们提出的研究将使用一种新的方法来识别利什曼尼亚生存能力涉及的关键参与者，从而使我们能够识别并创建高度特定的治疗剂，以破坏这些细胞过程。这种治疗方法可能会提供一种毒性较小，高度特定和有效的治疗利什曼病的方法。