Quantification of Differences in Downstream Signaling from Pathogenic FGFR2 Mutations Located in Different Functional Domains

Overview

Topic status: We're looking for students to study this topic.

The family of fibroblast growth factor (FGF) signalling molecules is complex with multiple FGF ligands and receptors. This family of receptors has been shown to be important in many aspects of development and disease especially cancer. Mutations in different domains of FGFR2 have been associated with germline skeletal syndromes and endometrial cancer. We hypothesize that these different mutations affect receptor localization and therefore activation of different downstream signalling pathways. Different activating mutations in FGFR2 have been shown to affect receptor conformation, trafficking to the cell surface and presumably signal predominantly from different subcellular compartments e.g. the cell membrane, ER and/or Golgi. We would like to develop a quantitative luciferase based assay to measuring the transcriptional activity of key effector proteins downstream of several different signalling pathways (e.g. MAPK, PI3K, Src, NFKb) by different pathogenic mutations in FGFR2. We have already developed a lot of the positive and negative constructs necessary for this project and believe this assay can also be adopted to answer a large number of additional unanswered questions in FGF/FGFR biology.

Hypothesis: Pathogenic mutations in FGFR2 differ in their subcellular localization and therefore differ in their quantitative and/or temporal activation of downstream signalling pathways to mediate different cellular phenotypes.

Aim 1: Establish stable cell lines expressing multiple different reporter constructs.

Aim 2: Validate the assay system using established activating and dominant negative mutations

Aim 3: Assess the effect of FGFR2 mutations located to the IgII-IgIII linker region, Ig III domain, juxtamembrane, transmembrane and kinase domains.

Aim 4: Correlate activation of signalling pathways with different cellular phenotypes

Methods and techniques that will be developed in the course of this project:

  • Cell culture (BaF3, endometrial epithelial cells)
  • Transfection and selection of stable cell lines
  • Intracellular signalling, Western blotting and luciferase assays
Study level
Honours
Supervisors
QUT
Organisational unit

Science and Engineering Faculty

Research area

Cell and Molecular Biosciences

Contact

Please contact the supervisor.