Functional Landscape of Glycosylation in Skin Cancer

项目总结/摘要 鳞状细胞癌是世界范围内的主要死亡原因。尽管癌症遗传学取得了进展， 在翻译后修饰中的作用知之甚少。蛋白质糖基化是最丰富的PTM， 在癌症中改变。聚糖不直接编码在基因组中，而是由多达10个 单糖以各种键合形成数千种结构，使其具有结构和功能 表征具有挑战性，并且在SCC中基本未知。为了解决这个问题，我建议使用鼠皮肤作为 高度易处理模型系统，以研究改变的糖基化、下游后果和鉴定新的 治疗在K99中，我将使用聚糖结合蛋白，代谢标记，先进的显微镜，MALDI 成像、糖组学、糖蛋白质组学和单细胞转录组学，以空间定位糖基化并识别 在遗传性、多次打击、组织学进展的SCC模型中， 乳头状瘤到鳞状细胞癌，化疗前和复发后（目的I）。接下来在K99中，我将使用我们的子宫 慢病毒技术，以覆盖表皮并进行大规模、无偏倚、体内功能基因组学 筛选约700个糖基因，以确定糖基化在SCC中的作用（Aim II）。最后在R 00中，我将联合收割机 糖蛋白组学，肿瘤和微环境的单细胞转录组学，蛋白质特异性聚糖编辑， 和高通量药物筛选，以确定聚糖驱动的肿瘤发生机制和新的治疗方法。 候选人（目标三）。我在糖生物学、医学肿瘤学方面的背景，以及来自领先的皮肤和 癌症生物学家Elaine Fuchs博士在洛克菲勒大学的优秀培训环境中， 纪念斯隆凯特琳癌症中心，作为一个博士后和医学肿瘤学研究员，将定位我， 解决这些重要的问题，学习技术，知识和领导力，以建立一个职业生涯， 在癌症糖生物学领域领先的独立研究者和科学家。

PROJECT SUMMARY/ABSTRACT Squamous cell carcinoma is a major cause of death worldwide. Despite advances in cancer genetics, alterations in post-translational modifications are poorly understood. Protein glycosylation is the most abundant PTM and altered in cancer. Glycans are not directly encoded in the genome but rather are comprised of up to ten monosaccharides in various linkages to form thousands of structures, making their structural and functional characterization challenging and largely unknown in SCC. To address this, I propose to use murine skin as a highly tractable model system to study altered glycosylation, downstream consequences, and identify novel therapies. In the K99, I will use glycan binding proteins, metabolic labeling, advanced microscopy, MALDI imaging, glycomics, glycoproteomics, and single cell transcriptomics to spatially map glycosylation and identify specific glycan and glycoprotein changes in a genetic, multi-hit, histologically progressive model of SCC from papilloma to SCC, before chemotherapy, and after recurrence (Aim I). Next in the K99, I will use our in utero lentivirus technology to transduce epidermis and perform a large-scale, unbiased, in vivo, functional genomics screen of the ~700 glycogenes to define the role of glycosylation in SCC (Aim II). Lastly in the R00, I will combine glycoproteomics, single cell transcriptomics of the tumor and microenvironment, protein-specific glycan editing, and high-throughput drug screens to identify mechanisms of glycan-driven tumorigenesis and novel therapeutic candidates (Aim III). My background in glycobiology, medical oncology, and mentorship from leading skin and cancer biologist Dr. Elaine Fuchs at the outstanding training environments at Rockefeller University and Memorial Sloan Kettering Cancer Center, as a postdoctoral and medical oncology fellow, will position me to tackle these important questions and learn techniques, knowledge, and leadership to establish a career as a leading independent investigator and physician-scientist in cancer glycobiology.