Investigating Membrane Trafficking Deficits in Choroideremia

研究无脉络膜血症的膜运输缺陷

• 批准号: 10282642
• 负责人: Abigail Fahim
• 金额: $ 23.76万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10282642
• 关键词: Address; Age related macular degeneration; Atrophic; Award; Biological Assay; Blindness; Blood Vessels; CRISPR/Cas technology; Cell Culture Techniques; Cell Line; Cell Proliferation; Cell Survival; Cells; Cessation of life; Childhood; Choroid; Choroideremia; Communication; Comparative Study; Defect; Degenerative Disorder; Derivation procedure; Development; Disease; Drug Targeting; Endothelial Cells; Equilibrium; Eye diseases; Functional disorder; Future; Genes; Goals; Growth Factor; Histopathology; In Vitro; Inherited; Investigation; Knowledge; Laboratories; Lead; Leadership; MT3 gene; Maintenance; Mass Spectrum Analysis; Mediating; Membrane; Membrane Lipids; Mentors; Mentorship; Modeling; Mutation; Pathologic; Pathway interactions; Patients; Phenotype; Photoreceptors; Professional Competence; Protein Secretion; Proteins; Rab escort protein; Reporting; Research Personnel; Resources; Retina; Retinal Degeneration; Retinal Diseases; Role; Signaling Molecule; Site; Small Interfering RNA; Structure of retinal pigment epithelium; Technical Expertise; Techniques; Testing; Therapeutic; Training; Vascular blood supply; Wallerian Degeneration; Work; career; career development; cell type; cytokine; drug development; experimental study; growth inhibitory proteins; high throughput screening; human stem cells; induced pluripotent stem cell; knock-down; monolayer; mouse model; novel; novel therapeutic intervention; overexpression; photoreceptor degeneration; prenylation; programs; protein transport; rab GTP-Binding Proteins; screening; skills; small molecule; stem cell biology; stem cells; targeted treatment; therapeutic target; trafficking; vascular bed; young adult

SUMMARY: Death of the choroid, the vascular bed underlying the retina, is a prominent pathologic hallmark of numerous inherited retinal degenerative diseases as well as age-related macular degeneration (AMD). Choroidal death is likely impacted by dysfunction of the retinal pigment epithelium (RPE), a monolayer of cells between the photoreceptors and the choroid. A critical knowledge gap impeding progress in therapeutic strategies is how RPE dysfunction contributes to choroidal death. Choroideremia is an inherited chorioretinal degeneration leading to early blindness with no current treatment. Several lines of evidence implicate the RPE as the primary site of degeneration, with secondary degeneration of the photoreceptors and choroid. The causative genetic defect is deficiency of CHM, encoding Rab escort protein 1 (REP-1), which facilitates prenylation of Rab GTPases, a critical step in membrane trafficking. RPE cells are highly polarized and differentially secrete numerous growth factors and signaling molecules in a directional manner towards the photoreceptors and choroid. The proposed studies will elucidate how RPE membrane trafficking defects in choroideremia alter protein secretion and how this impacts choroidal survival. The long-term goal of this work is to develop the necessary skills and expertise to establish an independent career focused on therapies targeting RPE dysfunction in retinal degenerative disease. The scientific objective is to understand mechanisms of choroidal death. We will test the hypothesis that alterations in RPE secretion of proteins in the vascular survival pathway lead to choroidal atrophy in choroideremia using a human stem cell derived RPE model. The career development objectives are to master stem cell culture and RPE derivation techniques, to develop expertise in membrane trafficking pathways of RPE, to become proficient in the techniques and principles of therapeutic target discovery, and to develop skills in leadership, mentorship, and scientific communication necessary to become a successful independent investigator. The proposed study will increase the candidate’s understanding of choroideremia and identify therapeutic targets in this untreatable blinding disease, which may also have broader implications for choroidal degeneration in AMD and other chorioretinal degenerative diseases.

摘要：视网膜下方的血管床的脉络膜死亡是一个突出的病理标志 许多遗传残留的退化性疾病以及与年龄相关的黄斑变性（AMD）。 脉络膜死亡可能受到视网膜色素上皮（RPE）功能障碍的影响，细胞的单层 在光感受器和脉络膜之间。关键的知识差距阻碍治疗的进步 策略是RPE功能障碍如何导致脉络膜死亡。脉络膜血症是一种遗传的脉络膜视网膜 变性导致早期失明，没有目前的治疗。几条证据暗示RPE 作为变性的主要部位，具有光感受器和脉络膜的继发性变性。这 致病遗传缺陷是CHM的缺陷，编码RAB伴随蛋白1（REP-1），促进 RAB GTPases的原始化，这是膜贩运的关键一步。 RPE细胞高度极化，并且 以方向方式向差异秘密的众多生长因子和信号分子朝向 感光体和脉络膜。拟议的研究将阐明RPE膜运输如何在 脉络膜血症改变蛋白质的分泌，以及这如何影响脉络膜存活。这项工作的长期目标 是发展必要的技能和专业知识，以建立专注于疗法的独立职业 靶向永久性退行性疾病中的RPE功能障碍。科学目标是了解 脉络膜死亡的机制。我们将检验以下假设：蛋白质中RPE分泌的改变 血管生存途径通过人类干细胞衍生的RPE导致脉络膜血症的脉络膜萎缩 模型。职业发展目标是掌握干细胞培养和RPE推导技术， 在RPE的膜贩运途径方面发展专业知识，以精通技术和 热目标发现的原则，并发展领导力，精通技术和科学技能 成为成功的独立调查员所必需的沟通。拟议的研究将增加 候选人对绒毛膜血症的理解并确定这种不可治疗的盲目性的治疗靶标 疾病，这可能对AMD和其他脉络膜结构的脉络膜变性具有更大的影响 退化性疾病。