Quantitative top-down proteomics of human colorectal cancer cells and tumors

项目摘要 据估计，有145,600人被诊断为结直肠癌(CRC)。 据预测，2019年有51,020人死于这种疾病 (https://seer.cancer.gov/statfacts/html/colorect.html).更好地了解CRC，请访问 分子水平肯定会带来更有效的治疗方法。基因组- 水平和转录组水平的信息不能准确地反映蛋白质水平 因为转录后调控可以调节蛋白质的表达和 因为翻译后修饰(PTM)可以影响蛋白质的功能。 结直肠癌的定量蛋白质组学研究是至关重要的。 许多自下而上的蛋白质组学研究已经在CRC细胞和肿瘤上完成，但 由于蛋白质序列较低，已获得的蛋白质形式信息有限 通常从自下而上的蛋白质组学获得的覆盖范围。不同的蛋白质形式 相同的基因可能具有截然不同的功能。我们假设大规模的 而人类结直肠癌细胞和肿瘤的定量自上而下蛋白质组学将提供 对CRC的新见解，导致更好的治疗方法。在这项提案中，我们将发展 新的分析工具，以提高自上而下的蛋白质组学的灵敏度和规模。新的 工具将使大规模和定量的自上而下的结直肠癌细胞蛋白质组学在 转移后以及来自林奇综合征患者的结直肠癌肿瘤。结果 从这项提案中得到的信息是极其重要的。新的分析工具将推动 自上而下的蛋白质组学的灵敏度提高了十倍，这将对 蛋白质组学社区用于大规模自上而下的大规模有限样本蛋白质组学。 大肠癌细胞转移前后的定量自上而下蛋白质组学研究 一种前所未有的资源，让癌症生物界获得新的见解 结直肠癌转移。林奇综合征组织的定量自上而下蛋白质组学将 阐明DNA错配修复基因突变和功能在慢性粒细胞白血病中的作用 蛋白形态水平的林奇综合征。

Project Summary It has been estimated that 145,600 people were diagnosed with colorectal cancer (CRC) and 51,020 deaths were predicted due to this disease in 2019 (https://seer.cancer.gov/statfacts/html/colorect.html). A better understanding of CRC at the molecular level will certainly lead to therapies that are more effective. The genome- level and transcriptome-level information cannot accurately reflect the protein-level information because post-transcriptional regulation can modulate protein expression and because post-translational modifications (PTMs) can influence protein function. Quantitative proteomic studies of CRC are vital. Many bottom-up proteomics studies have been completed on CRC cells and tumors, but limited information on proteoforms have been acquired due to low protein sequence coverages typically obtained from bottom-up proteomics. Different proteoforms from the same gene can have drastically different functions. We hypothesize that large-scale and quantitative top-down proteomics of human CRC cells and tumors will provide new insights into CRC, leading to better therapies. In this proposal, we will develop new analytical tools to boost the sensitivity and scale of top-down proteomics. The new tools will enable large-scale and quantitative top-down proteomics of CRC cells before and after metastasis as well as CRC tumors from patients with Lynch Syndrome. Results from this proposal are extremely important. The novel analytical tools will boost the sensitivity of top-down proteomics by tenfold and will be particularly useful for the proteomics community for large-scale top-down proteomics of mass-limited samples. Quantitative top-down proteomics of CRC cells before and after metastasis will generate an unprecedented resource for the cancer biology community to gain new insights into CRC metastasis. Quantitative top-down proteomics of the Lynch Syndrome tissues will elucidate the roles played by mutations and functions of DNA mismatch repair genes in Lynch Syndrome at the proteoform level.