Non-canonical NLRP3 Inflammasome Activation in Geographic Atrophy

地图样萎缩中的非典型 NLRP3 炎症小体激活

• 批准号: 9193873
• 负责人: Nagaraj Kerur
• 金额: $ 2.26万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9193873
• 关键词: Advisory Committees; Age related macular degeneration; Atrophic; Blindness; CASP1 gene; Caspase; Cell Culture Techniques; Cell Death; Cells; Cellular biology; Cessation of life; DICER1 gene; Dependency; Development; Disease; Educational workshop; Environment; Etiology; Eye; Faculty; Foundations; Goals; Grant; Health; Human; IL18 gene; Immune; Interferons; Interleukin-1; Interleukin-18; Investigation; Kentucky; Laboratories; Massachusetts; Measures; Mediating; Mentors; Modeling; Molecular; Monitor; Mus; Nature; Ophthalmology; Pathogenesis; Pathologic; Pathway interactions; Phase; Pilot Projects; Play; Proteomics; RNA; Regulation; Regulatory Pathway; Research; Research Personnel; Resources; Role; Scaffolding Protein; Signal Pathway; Signal Transduction; Structure of retinal pigment epithelium; Supervision; Therapeutic; Tissue Model; Training Activity; Transcript; Universities; Vision research; Work; Writing; age related; career; cytokine; geographic atrophy; human tissue; insight; macula; medical schools; member; mouse model; new therapeutic target; novel; programs; skills; vision science

DESCRIPTION (provided by applicant): This K99/R00 grant will facilitate the applicant's (Dr. Nagaraj Kerur) long-term goal to develop as an independent investigator in vision research. Geographic atrophy (GA) is an advanced form of age-related macular degeneration (AMD) characterized by central loss of vision due to retinal pigmented epithelium (RPE) degeneration. Currently there is no therapy available for GA because of its unclear etiology. We recently discovered that DICER1 deficit causes a pathologic accumulation of Alu RNA transcripts in the RPE of human GA eyes and that the accumulated Alu RNAs activate the NLRP3 inflammasome, which in turn leads to IL- 18/MyD88-dependent RPE cell death (Kaneko et al., Nature 2011; Tarallo et al., Cell 2012). In exciting, new preliminary studies we have discovered that Alu RNA-induced NLRP3 inflammasome activation occurs in a non-canonical fashion- a newly described pathway in which caspase 4 (aka caspase 11 in mouse) is required for the NLRP3 inflammasome activation (in contrast the canonical inflammasome activation is independent of caspase 4). The central goal of the proposed research is to define role of caspase 4-mediated non-canonical NLRP3 inflammasome activation in the context of GA and to identify critical signaling pathways regulating caspase 4 activation via the following aims: (1) Define the signaling pathways that regulate caspase 4 (aka caspase 11 in mouse) activation by Alu RNA; (2) Elucidate the mechanisms of caspase 4 (aka caspase 11 in mouse) activation of the NLRP3 inflammasome; and (3) Measure the activation of these non-canonical inflammasome signaling pathways in human GA. These studies will illuminate the molecular foundation of GA, unravel novel regulatory checkpoints of NLRP3 inflammasome activation, and provide a strong molecular foundation for exploiting new therapeutic targets in AMD. During the mentored phase, (K99) the applicant will carry out the proposed studies under direct supervision of the mentor Dr. Jayakrishna Ambati at the University of Kentucky. In addition, an advisory committee consisting of co-mentor, Dr. Katherine Fitzgerald and the faculty members at the Department of Ophthalmology and Visual Sciences will monitor the applicant's research progress and offer assistance in advancing the applicant's transition towards independent investigator. The Ambati Group at the University of Kentucky as well as the Fitzgerald Laboratory at the University of Massachusetts Medical School, both offer scientifically enriching environments and resources necessary for the advancement of the applicant's scientific career. The major focus of K99 phase will include: (1) mentored research to define molecular insights into the caspase 4 activation mechanism and to optimize experimental conditions for studies to be carried out in the R00 phase; and (2) training activities consisting of formal course work, seminars, workshops on grant writing and acquisition of other skill-sets required for successfully developing and leading an independent research program. During the R00 phase, the focus will be on: (1) deciphering the mechanisms by which caspase 4 regulates NLRP3 inflammasome activation; and (2) quantifying the activation of non-canonical inflammasome signaling pathways in human GA eyes. Overall this proposal will not only unravel the novel molecular mechanisms of RPE degeneration in GA, but will also equip the applicant with suitable skills to transition int an independent career in vision research.

描述（由申请人提供）：此K99/R00赠款将促进申请人（Nagaraj Kerur博士）的长期目标，以发展为视觉研究的独立研究者。地理萎缩（GA）是一种与年龄相关的黄斑变性（AMD）的高级形式，其特征是由于视网膜色素上皮（RPE）变性而导致视力丧失。目前，由于其病因不清楚，GA尚无治疗方法。我们最近发现，DICER1赤字会导致Alu RNA转录物在人GA眼中的病理积累，并且积累的Alu RNA激活了NLRP3炎性体，进而导致IL-18/MyD88依赖性RPE Cell死亡（Kaneko等，Kaneko等，自然2011; tarallo; tarallo; tarallo et al al an cyt an and cill al and cill al and and and and and and and and and and and and and and and。在令人兴奋的，新的初步研究中，我们发现Alu RNA诱导的NLRP3炎症体激活发生在非典型的时尚中 - 一种新描述的途径，其中caspase 4（鼠标中的AKA caspase 11中的aka caspase 11）需要用于NLRP3炎症体激活（相比之下，Canonical Inplampase implampase spivamasome激活了Caspase Activation Caspase 4）。拟议的研究的核心目的是在GA的背景下定义caspase 4介导的非传统NLRP3炎症体激活的作用，并确定通过以下目的来调节Caspase 4激活的关键信号通路：（1）定义调节caspase 4的信号通路（Alu caspase 4）（AKA Caspase 11 in Alu ru rnna by Alu rnna a Actration a Alu rnna caspase 11）; （2）阐明了NLRP3炎性体的激活caspase 4（aka caspase 11）的机制； （3）测量人类GA中这些非典型炎性体信号通路的激活。这些研究将阐明GA的分子基础，即NLRP3炎性体激活的新型调节检查点，并为利用AMD中新的治疗靶标提供了强大的分子基础。在指导阶段，（K99）申请人将在肯塔基大学的导师Jayakrishna Ambati博士的直接监督下进行拟议的研究。此外，由联合学者，凯瑟琳·菲茨杰拉德（Katherine Fitzgerald）博士和眼科科学系的教职员工组成的咨询委员会将监控申请人的研究进度，并为促进申请人向独立研究者的过渡提供帮助。肯塔基大学的Ambati集团以及马萨诸塞大学医学院的菲茨杰拉德实验室，都提供了科学丰富的环境和资源，以提高申请人的科学生涯。 K99阶段的主要重点将包括：（1）指导研究以将分子见解定义为caspase 4激活机制，并优化在R00阶段进行研究的实验条件； （2）包括正式课程工作，研讨会，授予写作研讨会以及成功制定和领导独立研究计划所需的其他技能集的研讨会。在R00阶段，将重点放在：（1）caspase 4调节NLRP3炎性体激活的机制； （2）量化人类GA眼中非典型炎性体信号通路的激活。总体而言，该提案不仅将揭示GA中RPE变性的新型分子机制，而且还将使申请人能够使申请人具有合适的技能，以转变视觉研究中的独立职业。