Discovering how oncogenes remodel the surfaceome of cells

标题：发现癌基因如何重塑细胞表面体 摘要：细胞表面是细胞与外界交流的主要枢纽。癌细胞 有无数的生存挑战，我们假设有许多 这些蛋白质从细胞表面开始，由大约3000-4000种蛋白质组成。 我们的主要目标是系统地了解癌细胞是如何 重塑它们的膜蛋白质组(表面体) 致癌转化以求生存。我们建议开发 使表面组学技术在人群、单细胞、 和组织水平。我们将生成基础数据集以 了解背后的分子逻辑和信号机制 驱动细胞转化的协调重塑事件。 (图1)。我们将重点研究突变激活的KRAS 和其他五个高增殖性癌基因(EGFR、BRAF、MEK、 AKT和PI3K)一起在近一半的人类中发现 癌症。我们将研究这些癌基因如何诱导协调 与同基因细胞相比，表型组的变化 类型，在患者来源的癌细胞和组织中。我们提出了新的 基于质谱学的方法，以允许量化 两种互补富集膜蛋白的方法 以表面多聚糖或表面N-末端α-胺为靶标。我们会 还开发了一种新的、高度敏感的多路传输技术 (噬菌体-抗体下一代测序，或PhaNGS)到 同时检测极小范围内的数百种表面蛋白 图1.(A)了解如何 KRAS等癌基因诱导 我们将进行大规模的细胞变革(B) 开发新技术以 测量表面体的变化 了解协调监管， 并确定新的药物靶点。 样本，甚至单个细胞。我领导的抗菌组中心，与重组人合作 抗体网络，利用噬菌体抗体产生了针对100种细胞表面蛋白的重组抗体 展示。每个Fab噬菌体本质上都是一种DNA条形码抗体，因为它显示了一个功能Fab 从含有编码DNA的噬菌体颗粒中分离出来。PhaNGS具有极大的潜力 多路、廉价和超灵敏的手段，探测数千个细胞或细胞系的变化 表面体。我很兴奋，这些研究将揭示如何协调膜蛋白的重塑 团队促成了细胞状态的改变。我相信我们还会发现新的生物标记物和抗癌药物 目标。

Title: Discovering how oncogenes remodel the surfaceome of cells Abstract: The cell surface is the primary hub for cells to communicate with the outside world. Cancer cells have numerous challenges for survival and we hypothesize many of these start at the cell surface made up of some 3000-4000 proteins. Our primary goal is to systematically understand how cancer cells remodel their membrane proteomes (surfaceomes) during oncogenic transformation to survive. We propose to develop enabling technologies for surfaceomics at the population, single cell, and tissue level. We will generate foundational data sets to understand the molecular logic and signaling mechanisms behind the coordinated remodeling events that drive cellular transformation. (Fig 1). We will focus our studies on mutationally activated KRAS and five other highly proliferative oncogenes (EGFR, BRAF, MEK, Akt and PI3K) that together are found in nearly half of all human cancers. We will study how these oncogenes induce coordinate changes in the surfaceome compared to isogenic cells of the same type, in patient derived cancer cells and tissues. We propose new mass spectrometry-based methods to allow for quantification of membrane proteins using two complementary enrichment methods that target surface glycans or surface N-terminal α-amines. We will also develop a new, highly sensitive and multiplexed technology (phage-antibody next generation sequencing, or PhaNGS) to simultaneously detect 100s of surface proteins in very small Figure 1. (A) To understand how oncogenes such as KRAS induce massive cellular changes we will (B) develop new technologies to measure surfaceome changes to understand coordinate regulation, and identify new drug targets. samples or even single cells. The Antibiome Center, which I direct, in collaboration with the Recombinant Antibody Network, has produced recombinant antibodies to 100s of cell surface proteins using Fab-phage display. Each Fab-phage is essentially a DNA barcoded antibody because it has a functional Fab displayed from the phage particle with the DNA encoding it inside. PhaNGS has tremendous potential for highly multiplexed, inexpensive and ultrasensitive means of probing 1000s of cells or cell lines for changes in their surfaceomes. I am excited that these studies will reveal how coordinate remodeling of membrane protein teams contributes to cell state changes. I believe we will also discover new biomarkers and cancer drug targets.