Modeling Retinoblastoma Initiation Using 3D-Retinal Organoids

项目摘要 癌症遗传学的几个里程碑式的发现来自于对一种罕见的儿童癌症的研究， 称为视网膜母细胞瘤。与此同时，临床前测试和临床研究的进展导致了 改善患有这种毁灭性疾病的儿童的预后。尽管取得了这些进展， 视网膜母细胞瘤领域的基本问题仍然没有答案。视网膜细胞是否完全转化 一旦它们维持RB1基因的双等位基因失活或视网膜母细胞瘤肿瘤发生是多阶段的， 过程？为什么一些具有相同生殖系RB1突变的家庭成员有双侧多灶性 视网膜母细胞瘤在年轻的时候，而其他人没有疾病的证据？治疗是否会导致 肿瘤细胞克隆进化和选择导致肿瘤进展和去核？这些问题 因为视网膜母细胞瘤不做活检，只做眼球摘除术， 对于高级阶段的眼睛。为了克服该领域的这一障碍，我们开发了第一个 使用从患有视网膜母细胞瘤的患者产生的3D视网膜类器官的自发性人视网膜母细胞瘤肿瘤模型 生殖系RB1突变。我组建了一个具有计算和STEM专业知识的多学科团队 细胞生物学，肿瘤学，病理学，流行病学和生物统计学，使用这种创新的新模式， 视网膜母细胞瘤回答3个具体目标的基本问题。我们将确定视网膜母细胞瘤 如果分子、细胞或遗传因素有助于 差异的表达率和表达（目标2），如果有克隆选择与治疗（目标3）。我有 在视网膜母细胞瘤遗传学和将基础科学发现转化为临床试验方面有着良好的记录。这 一项提案将通过一种新的维持疗法的临床前测试来影响视网膜母细胞瘤患者 (Aim 3）在完成治疗后的最初几个月内，防止新的肿瘤在周边视网膜中形成。 化疗它也可能有助于识别具有生殖系RB1突变的视网膜母细胞瘤幸存者的子集 具有发展第二种恶性肿瘤的超高风险，需要更广泛的癌症筛查（Aim 2）。没有其他中心有团队、资源、专业知识或工具来执行所提出的研究 并有效地将最有希望的发现直接转移到临床试验中。

PROJECT SUMMARY Several landmark discoveries in cancer genetics have come from studies on a rare childhood cancer of the developing retina called retinoblastoma. In parallel, advances in preclinical testing and clinical research has led to improvements in outcome for children with this devastating disease. Despite these advances, there are still fundamental questions in the retinoblastoma field that remain unanswered. Are retinal cells fully transformed once they sustain biallelic inactivation of the RB1 gene or is retinoblastoma tumorigenesis a multistage process? Why do some family members with the same germline RB1 mutation have bilateral multifocal retinoblastoma at a young age while others have no evidence of disease? Can treatment induce a process of tumor cell clonal evolution and selection that leads to tumor progression and enucleation? These questions have been impossible to answer because retinoblastomas are not biopsied and enucleation is only performed for advanced stage eyes. In order to overcome this barrier in the field, we have developed the first spontaneous human retinoblastoma tumor model using 3D retinal organoids produced from patients with germline RB1 mutations. I have assembled a multidisciplinary team with expertise in computational and stem cell biology, oncology, pathology, epidemiology and biostatistics to use this innovative new model of retinoblastoma to answer fundamental questions in 3 specific aims. We will determine if retinoblastoma progresses through a multistep process (Aim 1), if molecular, cellular or genetic factors contributes to differences in penetrance and expressivity (Aim 2) and if there is clonal selection with treatment (Aim 3). I have a proven record in retinoblastoma genetics and of moving basic science discoveries into clinical trials. This proposal will impact patients with retinoblastoma through preclinical testing of a novel maintenance therapy (Aim 3) to prevent new tumors from forming in the peripheral retina in the first few months after completion of chemotherapy. It may also help to identify a subset of retinoblastoma survivors with germline RB1 mutations that have an ultra-high risk of developing a 2nd malignancy and require more extensive cancer screening (Aim 2). No other center has the team, resources, expertise, or tools available to perform the studies presented here and efficiently move the most promising findings directly into a clinical trial.