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

项目摘要 据估计，145，600人被诊断患有结直肠癌（CRC） 预计2019年将有51，020人死于这种疾病 （https：//seer.cancer.gov/statfacts/html/colorect.html）。更好地了解CRC， 分子水平肯定会导致更有效的治疗。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 因为转录后调节可以调节蛋白质表达， 因为翻译后修饰（PTMs）可以影响蛋白质功能。 CRC的定量蛋白质组学研究至关重要。 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 由于蛋白质序列低，获得的蛋白质型信息有限 通常从自下而上的蛋白质组学获得的覆盖率。不同的蛋白形式 同样的基因可以有截然不同的功能。我们假设大规模的 人类CRC细胞和肿瘤的定量自上而下的蛋白质组学将提供 对CRC的新见解，导致更好的治疗。在本提案中，我们将开发 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 工具将使CRC细胞的大规模和定量自上而下的蛋白质组学， 和转移后以及来自Lynch综合征患者的CRC肿瘤。结果 这一提议非常重要。新的分析工具将促进 自上而下的蛋白质组学的灵敏度提高了十倍，这将特别适用于 蛋白质组学社区，用于对质量有限的样品进行大规模自上而下的蛋白质组学研究。 CRC细胞转移前后的定量自上而下的蛋白质组学将产生 癌症生物学社区获得新见解的前所未有的资源， CRC转移。林奇综合征组织的定量自上而下的蛋白质组学将 阐明DNA错配修复基因的突变和功能在 蛋白质水平的林奇综合征。

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.