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Discovering how oncogenes remodel the surfaceome of cells

发现癌基因如何重塑细胞表面组

基本信息

批准号：
10212408
负责人：
JAMES A WELLS
金额：
$ 53.49万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2023-06-30
项目状态：
已结题

项目摘要

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.

原标题：发现癌基因如何重塑细胞表面翻译后摘要：细胞表面是细胞与外界沟通的主要枢纽。癌细胞有许多生存的挑战，我们假设许多这些起始于由大约3000-4000种蛋白质组成的细胞表面。我们的主要目标是系统地了解癌细胞重塑他们的膜蛋白质组（表面组），致癌转化才能生存。我们建议发展使表面组学技术能够在群体，单细胞，和组织水平。我们将生成基础数据集，理解背后的分子逻辑和信号机制协调的重塑事件驱动细胞转化。 (Fig 1）。我们将把研究重点放在突变激活的KRAS上，和五种其它高度增殖的癌基因（EGFR，BRAF，MEK， Akt和PI 3 K），它们共同存在于近一半的人类中，癌的我们将研究这些癌基因如何诱导协调与同基因细胞相比，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的方法，以允许定量使用两种互补富集方法的膜蛋白靶向表面聚糖或表面N-末端α-胺。我们将我还开发了一种新的，高灵敏度和多路复用技术，（噬菌体抗体下一代测序，或PhaNGS），在非常小的空间内同时检测数百种表面蛋白图1. (A)了解如何致癌基因如KRAS诱导我们将看到大量的细胞变化（B）开发新技术以测量表面的变化，了解协调调节，并确定新的药物靶点。甚至是单个细胞。我领导的抗生素组中心与重组抗体网络，已经使用Fab噬菌体产生了针对100多种细胞表面蛋白的重组抗体显示.每个Fab噬菌体基本上是DNA条形码化抗体，因为它具有展示的功能性Fab，从含有编码它的DNA的噬菌体颗粒中分离出来。PhaNGS具有巨大的潜力，多路复用的，廉价的和超灵敏的手段探测1000个细胞或细胞系的变化，表面生物我很高兴这些研究将揭示如何协调重塑膜蛋白团队有助于细胞状态的变化。我相信我们还将发现新的生物标志物和抗癌药物目标的