Leveraging multi-omics to define the role of epigenetic regulation by PRC2 in Esthesioneuroblastoma

摘要 永久性嗅觉丧失是一种与嗅觉神经母细胞瘤(ENB)相关的常见疾病，这种肿瘤被认为是 起源于鼻子里的嗅觉上皮。这是由于需要进行大量手术切除，而不考虑 肿瘤分级。因此，开发新的医疗治疗策略对于保护嗅觉是必要的 在这群病人中。嗅觉上皮是一个神经性的小生境，它产生神经元和支持 整个生命中的上皮细胞，以及各种未分化的正常上皮细胞增殖和 在eNB中观察到了发育情况。表观遗传调节因子多梳抑制复合体2(PRC2)具有 被牵连到推动全身干细胞壁龛中的增殖细胞状态。我们已经证明了 在嗅觉上皮细胞中，PrC2对基底细胞的增殖起着至关重要的作用，我们已经证实它被表达 在eNb的增殖细胞中。由于ENB是一种罕见的肿瘤，机械性的生物学研究很少，而且没有细胞 线条或鼠标模型存在。此外，大多数eNBs并不共享共同的驱动突变， 强调表观遗传调控的重要性。在这里，我建议利用新开发的多组体 研究人类eNB中PRC2的表达如何指定细胞状态并与肿瘤有关 成长。我将对多个肿瘤进行单细胞染色质和转录分析，以确定 染色体可及性。此外，我将在人类eNB样本中使用药理学检测来评估 下游对转录的影响。最后，空间转录组学将被用来定义eNB细胞和 分子组成，包括PRC相关的表达，以及与正常嗅觉的比较 上皮细胞，以阐明eNB内肿瘤和免疫细胞之间的细胞信号动力学 微环境。拟议的研究完成后，将确定eNB与 目标是确定新的可用药目标，从而提高存活率和气味保存率。总而言之， 本提案中概述的研究和培训计划与布拉德利博士的专家指导相结合 戈尔茨坦将为我作为外科科学家的成功职业生涯所需的严格培训做好准备 鼻科和前颅底外科。

ABSTRACT Permanent smell loss is a common morbidity associated with esthesioneuroblastoma (ENB), a tumor thought to arise from the olfactory epithelium in the nose. This is due to necessity for bulk surgical resection, regardless of tumor grade. As such, the development of novel medical treatment strategies is necessary to preserve olfaction in this patient population. The olfactory epithelium is a neurogenic niche that produces neurons and supporting epithelial cells throughout life, and various undifferentiated stages of normal epithelial proliferation and development have been observed in ENB. The epigenetic regulator polycomb repressive complex 2 (PRC2) has been implicated in driving proliferative cell states in stem cell niches throughout the body. We have shown that PRC2 is crucial for basal cell proliferation in the olfactory epithelium, and we have identified that it is expressed in proliferating cells in ENB. Because ENB is a rare tumor, mechanistic biological studies are sparse, and no cell lines or mouse models exist. Furthermore, the majority of ENBs do not share common driver mutations, emphasizing the importance of epigenetic regulation. Here I propose to utilize newly-developed multi-omic studies to investigate how PRC2 expression in human ENB specifies cellular states and contributes to tumor growth. I will perform single cell chromatin and transcriptomic assays on multiple tumors to identify areas of chromosomal accessibility. Furthermore, I will use pharmacologic assays in human ENB samples to assess the downstream effects on transcription. Finally, spatial transcriptomics will be used to define ENB cellular and molecular composition, including PRC-associated expression, and how this compares to normal olfactory epithelium, to elucidate cellular signaling dynamics among tumor and immune cells within the ENB microenvironment. Completion of the proposed studies will define epigenetic drivers co-opted by ENB with the goal of identifying new druggable targets, allowing for improved survival and preservation of smell. Altogether, the research and training plan outlined in this proposal combined with expert mentorship from Dr. Bradley Goldstein will prepare me with the rigorous training necessary for a successful career as a surgeon-scientist in rhinology and anterior skull base surgery.