Discovery of immunotherapeutic biologics against the RAS-driven extracellular proteome

 描述(申请人提供)：这项建议的目标是(I)了解RAS癌基因转化如何改变细胞表面蛋白质组，以及(Ii)产生针对RAS驱动的癌细胞细胞外蛋白质组的重组抗体，作为诊断生物标记物和潜在的治疗试剂。RAS是最常见的癌基因，在超过30%的癌症中发生突变。正常细胞向恶性状态的转变与蛋白质组的许多变化有关，包括细胞表面的蛋白质，以及细胞形态的变化，并导致信号异常和增殖。识别RAS驱动的癌细胞表面的细胞外蛋白质组变化将有助于更好地理解导致这些表型变化的潜在机制。将通过几种方法研究K-RAS突变引起的变化：(I)与正常细胞相比，确定RAS诱导的细胞外表面差异表达的蛋白质靶点；(Ii)针对已确定的胞外靶点产生抗体；以及(Iii)评估针对患者来源的癌细胞株和组织的抗体的治疗潜力。因此，这项提议的目的是利用新的膜蛋白质组学和新的免疫治疗技术创建一个理解和询问致癌细胞表面的通用平台，最终可以扩展到许多其他癌症。这些数据集、技术和分子工具包的开发将为癌基因转化的细胞如何重塑其表面提供新的见解。

 DESCRIPTION (provided by applicant): The goals of this proposal are to (i) understand how Ras oncogene transformation changes the cell surface proteome and to (ii) generate recombinant antibodies against the extracellular proteome of Ras-driven cancer cells to serve as diagnostic biomarkers and potential therapeutic reagents. Ras is the most common oncogene and is mutated in over 30% of cancers. Transformation of a normal cell to a malignant state is associated with many changes in the proteome including proteins at the cell surface, as well as changes in cell morphology, and result in aberrant signaling and proliferation. Identification of extracellular proteome changes at the surface of Ras-driven cancer cells will lead to a better understanding of the underlying mechanisms that cause these phenotypic alterations. The changes induced by K-Ras mutants will be investigated using several approaches: (i) determination of the differentially expressed protein targets on the extracellular surface of the Ras-induced cells in comparison to normal cells, (ii) antibody generation against the identified extracellular targets, and (iii) assessment of the therapeutic potential of the antibodies against patient-derived cancer cell lines and tissues. Thus, this proposal will be aimed at creating a general platform for understanding and interrogating oncogenic cellular surfaces with novel membrane proteomics and new immunotherapeutic technologies, which can ultimately be extended to many other cancers. The development of these data sets, technologies, and molecular toolkits will provide new insights into how oncogene transformed cells remodel their surfaces.