3-Dimensional Retinal Organoid Platform for the Study of Retinoblastoma

用于视网膜母细胞瘤研究的 3 维视网膜类器官平台

• 批准号: 10263901
• 负责人: JAMES WILLIAM HARBOUR
• 金额: $ 49.28万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10263901
• 关键词: 3-Dimensional; Address; Affect; Animal Model; Animals; Bar Codes; Biocompatible Materials; Biological; Biological Models; Biomimetics; Bioreactors; CRISPR/Cas technology; Cells; Child; Clinical Management; Clone Cells; Culture Techniques; Custom; DNA sequencing; Data; Defect; Dependence; Developed Countries; Developing Countries; Development; Disease; Drug Targeting; Embryo; Engineering; Environment; Equilibrium; Event; Excision; Eye; Genes; Genetic; Genetic Transcription; Genetically Engineered Mouse; Health; Human; Hypoxia; Invaded; Knowledge; Laboratories; Lead; Left; Life; Light; Malignant - descriptor; Malignant Childhood Neoplasm; Malignant Neoplasms; Modeling; Molecular; Molecular Abnormality; Mus; Mutation; Nature; Neoplasm Metastasis; Neural Retina; Normal tissue morphology; Nuclear Orphan Receptor; Optic Nerve; Organ; Organoids; Output; Oxygen; Parents; Pathway interactions; Patients; Pattern; Pharmacology; Phase; Phenotype; Photoreceptors; Physiological; Procedures; Proliferating; RB1 gene; Resolution; Retina; Retinal Cone; Retinal Neoplasms; Retinal Photoreceptors; Retinoblastoma; Rod; Savings; Seeds; Stimulus; Survival Rate; System; TP53 gene; Technology; Testing; Therapeutic; Time; Tissue Engineering; Tissues; Tumor Suppressor Proteins; Undifferentiated; Vision; Visual; Visual impairment; base; bioinformatics pipeline; cancer genomics; cell type; childhood cancer mortality; clinical phenotype; cost; design; differentiation protocol; drug development; druggable target; estrogen-related receptor; genetic analysis; genetic corepressor; genetic manipulation; genomic aberrations; human disease; improved; in silico; in utero; induced pluripotent stem cell; malignant neoplasm of eye; mouse model; multidisciplinary; mutant; neoplastic cell; new technology; next generation; p107 protein; premalignant; preservation; psychologic; retinal neuron; retinal progenitor cell; retinogenesis; single-cell RNA sequencing; social stigma; stem cell biology; targeted treatment; therapeutic candidate; tissue culture; transcriptome sequencing; tumor; tumor progression

Project Summary/Abstract Retinoblastoma (Rb) is the most common eye cancer in children and a significant contributor to childhood cancer deaths worldwide. Rb tumors arise from biallelic loss of the RB1 gene during retinal development in-utero, and are uniformly lethal if left untreated. While clinical management of the tumor has advanced in recent decades, the non-specific and highly toxic nature of current therapies frequently result in life-long visual compromise and health complications. Similarly, the exorbitant cost and limited access to advanced procedures render the best treatment options inaccessible to most children worldwide, where eye removal remains the only life-saving option for patients. However, the social stigma and psychological effects of eye removal often lead parents to forego treatment, resulting in survival rates below 50% in many regions of the world. The most significant barrier to progress in developing targeted selective therapies for retinoblastoma is the gap in knowledge regarding the molecular drivers of retinoblastoma progression. Currently, no laboratory models exist, animal or otherwise, which faithfully recapitulate the human manifestation of this disease, and the limited amount of information available comes from tumors which necessitate removal, thus only providing a snapshot of the molecular mechanisms present in advanced cases. The ability to study these tumors in earlier stages, including their pre- malignant phase would shed light into the molecular drivers of Rb progression, lead to more targeted therapies which obviate the need for eye removal, ideally preserving vision but, more importantly, save lives. Recently, we have developed a human iPSC-based 3-Dimensional retinal organoid differentiation protocol and tissue culture bioreactor system in which we can control various environmental parameters critical for proper retinal development. Using this platform, we can use gene editing to introduce the most prevalent mutations found in Rb tumors and evaluate their effect on retinal development and tumor formation. We have shown how some of these mutations lead to amplification of a pre-malignant undifferentiated mutant cell clone, and disrupt terminal differentiation of other retinal cell types. We now propose to establish this platform as the first model system in which the various aspects of human Rb tumors can be elucidated, from their initiation through the drivers of malignant transformation. In order to achieve this, we aim to 1) Optimize, validate, and deploy our tissue bioreactor system to recapitulate the microenvironment of the developing retina, 2) use gene editing technology to establish the physiologic pattern of mutation acquisition during retinogenesis which leads to Rb tumor formation, and 3) use our 3D retinal organoid platform to screen pharmacological compounds that can address the molecular determinants of Rb tumor progression.

项目总结/摘要 视网膜母细胞瘤（Rb）是儿童最常见的眼癌，也是儿童癌症的重要贡献者 全球死亡。Rb肿瘤是由于视网膜在子宫内发育过程中RB 1基因的双等位基因缺失引起的， 如果不治疗的话都是致命的虽然近几十年来肿瘤的临床治疗取得了进展， 目前治疗非特异性和高毒性的性质经常导致终身视力损害， 健康并发症。同样，高昂的费用和获得先进程序的机会有限， 全球大多数儿童无法获得治疗选择，摘除眼睛仍然是唯一挽救生命的选择 对患者然而，摘除眼睛的社会耻辱和心理影响往往导致父母放弃 在世界许多地区，存活率低于50%。最大的障碍是 视网膜母细胞瘤的靶向选择性治疗的进展是关于视网膜母细胞瘤的知识的差距。 视网膜母细胞瘤进展的分子驱动因素。目前，没有实验室模型存在，动物或其他， 它忠实地概括了这种疾病的人类表现，以及有限的信息量， 可用的来自需要切除的肿瘤，因此仅提供了分子水平的快照。 在先进的情况下存在的机制。在早期阶段研究这些肿瘤的能力，包括它们的前 恶性阶段将揭示Rb进展的分子驱动因素，导致更多的靶向治疗 这消除了摘除眼睛的需要，理想地保留了视力，但更重要的是，可以挽救生命。最近我们 已经开发了基于人iPSC的三维视网膜类器官分化方案和组织培养 生物反应器系统，我们可以控制各种环境参数的关键适当的视网膜 发展使用这个平台，我们可以使用基因编辑来引入在基因组中发现的最常见的突变。 Rb肿瘤，并评估其对视网膜发育和肿瘤形成的影响。我们已经展示了一些 这些突变导致癌前未分化突变细胞克隆的扩增，并破坏终末 其他视网膜细胞类型的分化。我们现在建议建立这个平台，作为 其中人类Rb肿瘤的各个方面可以阐明，从它们的开始通过驱动程序， 恶性转化为了实现这一目标，我们的目标是：1）优化、验证和部署我们的组织 生物反应器系统来重现发育中的视网膜的微环境，2）使用基因编辑技术 建立视网膜发生过程中突变获得的生理模式，从而导致Rb肿瘤 形成，以及3）使用我们的3D视网膜类器官平台来筛选可以解决 Rb肿瘤进展的分子决定因素。