Determining the developmental origins of retinoblastoma

确定视网膜母细胞瘤的发育起源

• 批准号: 9891844
• 负责人: Jacqueline Norrie
• 金额: $ 6.74万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9891844
• 关键词: 2 year old; Address; Affect; Age; Alleles; Animal Model; Biological Markers; Biological Models; Caring; Cell Line; Cells; Child; Childhood; Data; Development; Diagnosis; Disease; Disease model; Electron Microscopy; Excision; Eye; Eye Neoplasms; Genetic; Genetic Predisposition to Disease; Germ-Line Mutation; Goals; Human; Immune; Inherited; Life; Loss of Heterozygosity; Malignant Neoplasms; Modeling; Molecular; Mus; Mutation; Oncogenic; Organoids; Parents; Patients; Pattern; Play; Preclinical Testing; Protocols documentation; Quality of life; RB1 Gene Inactivation; RB1 gene; Reporter; Reporting; Retina; Retinoblastoma; Role; SYK gene; Specificity; Structure; Study models; Survival Rate; System; Time; Tumor stage; Untranslated RNA; Xenograft procedure; base; cancer initiation; cell type; chemotherapy; effective therapy; exome sequencing; functional loss; human embryonic stem cell; human model; in vivo; induced pluripotent stem cell; insight; malignant neoplasm of eye; mouse model; novel; outcome forecast; research clinical testing; single cell sequencing; targeted treatment; trait; transcriptome sequencing; tumor; tumor growth; tumor initiation; tumorigenesis; virtual

Project Summary/Abstract Retinoblastoma is the most common pediatric eye cancer and is caused by the biallelic inactivation of the tumor suppressor gene RB1. There are between 200 and 300 new patients with retinoblastoma in the U.S. each year, most diagnosed before two years of age. When caught early, and with proper care, survival from retinoblastoma is relatively high. However, treatments, including enucleations, or removal of the eye, have lasting effects on the lives of the children. Furthermore, retinoblastomas with extraocular involvement have a much poorer prognosis. While animal models of retinoblastoma have been useful for preclinical and clinical testing of retinoblastoma treatment, mice with loss of RB1 never develop retinoblastoma and therefore cannot accurately recapitulate human retinoblastoma formation. This has led to gaps in our understanding of the tumor initiation, development, and cellular specificity of human retinoblastoma. The goal of this study is to use our new model of spontaneous human retinoblastoma using patient derived iPSCs to study the developmental origin(s) of retinoblastoma and characterize the mechanisms of the second hit mutation. We have generated 15 induced pluripotent stem cell (iPSC) lines derived from patients with germline mutations in RB1. Using an optimized retinal differentiation culture, we have differentiated the patient lines into retina, dissociated the organoids, injected the cells into mice to screen for tumor formation. The resulting tumors have expression patterns similar to retinoblastoma patient derived xenografts. I now have a model system to study retinoblastoma tumor formation during retinal differentiation. I will use our new retinoblastoma reporter line in H9 hESCs, SYK-GFP, and a combination of immunostaining, single cell sequencing and electron microscopy to determine cell of origin or retinoblastoma. Additionally, I will analyze the tumors over multiple rounds of differentiation to determine the mechanism of the second hit and if there is any genetic predisposition to the loss of heterozygosity that occurs in patients with germline RB1 mutations. This study will allow us, for the first time, to answer question about retinoblastoma initiation and progression. Defining the cell of origin or retinoblastoma and the mechanism of the second hit of RB1 will not only increase our depth of understanding of retinoblastoma development but can be more generally applied to the fundamental understanding of cancer initiation. Furthermore, the data provided by this study will hopefully provide insight to create more targeted therapies for children with retinoblastoma.

项目总结/摘要 视网膜母细胞瘤是最常见的儿童眼癌，是由视网膜母细胞瘤基因的双等位基因失活引起的。 肿瘤抑制基因RB 1。在美国有200到300名新的视网膜母细胞瘤患者。 大多数在两岁之前被诊断出来。如果及早发现，并给予适当的照顾， 视网膜母细胞瘤的发病率相对较高。然而，治疗，包括摘除或摘除眼睛， 对孩子们的生活产生持久的影响。此外，视网膜母细胞瘤伴眼外受累， 预后差得多。虽然视网膜母细胞瘤的动物模型对于临床前和临床研究是有用的， 在视网膜母细胞瘤治疗的测试中，RB 1缺失的小鼠从未发生视网膜母细胞瘤，因此不能 准确地再现了人类视网膜母细胞瘤的形成。这导致了我们在理解 人视网膜母细胞瘤的肿瘤发生、发展和细胞特异性。 本研究的目的是使用我们的新的自发性人视网膜母细胞瘤模型， iPSCs研究视网膜母细胞瘤的发育起源，并表征第二次视网膜母细胞瘤的机制。 命中突变。我们已经产生了15个诱导多能干细胞（iPSC）系，其来源于患有以下疾病的患者： RB 1的生殖细胞突变。使用优化的视网膜分化培养，我们已经区分了病人 将细胞线植入视网膜，解离类器官，将细胞注射到小鼠体内以筛选肿瘤形成。的 所得肿瘤具有与视网膜母细胞瘤患者来源的异种移植物相似的表达模式。我现在有一个 模型系统研究视网膜分化过程中视网膜母细胞瘤肿瘤形成。我会用我们新的 H9 hESC中的视网膜母细胞瘤报告细胞系、SYK-GFP和免疫染色、单细胞 测序和电子显微镜以确定细胞来源或视网膜母细胞瘤。此外，我将分析 肿瘤在多轮分化，以确定第二次打击的机制，如果有 生殖系RB 1突变患者中发生杂合性丢失的任何遗传倾向。 这项研究将使我们首次回答有关视网膜母细胞瘤发生和进展的问题。 确定视网膜母细胞瘤的起源细胞和RB 1第二次击中的机制不仅会增加 我们对视网膜母细胞瘤发展的理解深度，但可以更普遍地应用于 对癌症起源的基本认识。此外，本研究提供的数据有望 为视网膜母细胞瘤儿童创造更多靶向治疗提供见解。