QI-FRET: a new tool in Receptor Tyrosine Kinase research

描述（由申请人提供）：QI-FRET：受体酪氨酸激酶研究中的一种新工具Kalina Hristova，Johns霍普金斯大学受体酪氨酸激酶（RTK）的配体非依赖性同源和异源二聚化是一种尽管与各种人类病理学有关但尚未完全理解的过程。知识的缺乏主要是由于缺乏适当的方法来测量全长RTK的二聚化能量学。知识的缺乏反过来又是开发有效RTK靶向疗法的瓶颈。在这里，我们建议（i）开发一种定量成像FRET（QI-FRET）的方法，产生二聚自由能在质膜衍生的囊泡和（ii）表征的同源和异源二聚自由能的ErbB和FGF受体，选择，因为他们非常强大的联系到人类疾病。这项工作完成后，我们将能够预测配体独立的二聚化的程度，从而生物活性，作为RTK表达的函数。将建立的方法和获得的知识将最终有助于寻找和完善有效的RTK靶向治疗癌症和生长障碍。 公共卫生相关性：我们建议（i）开发一种定量成像FRET（QI-FRET）方法，产生质膜衍生囊泡中的二聚化自由能，并（ii）表征ErbB和FGF受体的同源和异源二聚化自由能，选择这些受体是因为它们与人类疾病有很强的联系。这项工作完成后，我们将能够预测配体独立的二聚化的程度，从而生物活性，作为RTK表达的函数。将建立的方法和获得的知识将最终有助于寻找和完善有效的RTK靶向治疗癌症和生长障碍。

DESCRIPTION (provided by applicant): QI-FRET: a new tool in Receptor Tyrosine Kinase research Kalina Hristova, Johns Hopkins University Ligand-independent homo and heterodimerization of receptor tyrosine kinases (RTKs) is a process that is not well understood despite being implicated in various human pathologies. The lack of knowledge is mainly due to a lack of an appropriate methodology to measure the dimerization energetics of full-length RTK. The lack of knowledge is, in turn, a bottleneck in developing effective RTK-targeted therapies. Here we propose to (i) develop a quantitative imaging FRET (QI-FRET) method that yields dimerization free energies in plasma membrane-derived vesicles and to (ii) characterize the homo and heterodimerization free energies of the ErbB and FGF receptors, chosen because of their very strong link to human disease. Upon the completion of this work, we will be able to predict the degree of ligand-independent dimerization, and thus biological activity, as a function of RTK expression. The method that will be established and the knowledge gained will ultimately aid the search and refinement of effective RTK-targeted treatments for cancers and growth disorders. PUBLIC HEALTH RELEVANCE: We propose to (i) develop a quantitative imaging FRET (QI-FRET) method that yields dimerization free energies in plasma membrane-derived vesicles and to (ii) characterize the homo and heterodimerization free energies of the ErbB and FGF receptors, chosen because of their very strong link to human disease. Upon the completion of this work, we will be able to predict the degree of ligand-independent dimerization, and thus biological activity, as a function of RTK expression. The method that will be established and the knowledge gained will ultimately aid the search and refinement of effective RTK-targeted treatments for cancers and growth disorders.