Reference - Detail
| RRC ID | 79173 |
|---|---|
| Author | Fasano G, Petrini S, Bonavolontà V, Paradisi G, Pedalino C, Tartaglia M, Lauri A. |
| Title | Assessment of the FRET-based Teen sensor to monitor ERK activation changes preceding morphological defects in a RASopathy zebrafish model and phenotypic rescue by MEK inhibitor. |
| Journal | Mol Med |
| Abstract |
BACKGROUND:RASopathies are genetic syndromes affecting development and having variable cancer predisposition. These disorders are clinically related and are caused by germline mutations affecting key players and regulators of the RAS-MAPK signaling pathway generally leading to an upregulated ERK activity. Gain-of-function (GOF) mutations in PTPN11, encoding SHP2, a cytosolic protein tyrosine phosphatase positively controlling RAS function, underlie approximately 50% of Noonan syndromes (NS), the most common RASopathy. A different class of these activating mutations occurs as somatic events in childhood leukemias. METHOD:Here, we evaluated the application of a FRET-based zebrafish ERK reporter, Teen, and used quantitative FRET protocols to monitor non-physiological RASopathy-associated changes in ERK activation. In a multi-level experimental workflow, we tested the suitability of the Teen reporter to detect pan-embryo ERK activity correlates of morphometric alterations driven by the NS-causing Shp2D61G allele. RESULTS:Spectral unmixing- and acceptor photobleaching (AB)-FRET analyses captured pathological ERK activity preceding the manifestation of quantifiable body axes defects, a morphological pillar used to test the strength of SHP2 GoF mutations. Last, the work shows that by multi-modal FRET analysis, we can quantitatively trace back the modulation of ERK phosphorylation obtained by low-dose MEK inhibitor treatment to early development, before the onset of morphological defects. CONCLUSION:This work proves the usefulness of FRET imaging protocols on both live and fixed Teen ERK reporter fish to readily monitor and quantify pharmacologically- and genetically-induced ERK activity modulations in early embryos, representing a useful tool in pre-clinical applications targeting RAS-MAPK signaling. |
| Volume | 30(1) |
| Pages | 47 |
| Published | 2024-4-9 |
| DOI | 10.1186/s10020-024-00807-w |
| PII | 10.1186/s10020-024-00807-w |
| PMID | 38594640 |
| PMC | PMC11005195 |
| MeSH | Adolescent Animals Fluorescence Resonance Energy Transfer Humans Mitogen-Activated Protein Kinase Kinases / genetics Mitogen-Activated Protein Kinase Kinases / metabolism Mutation Noonan Syndrome* / genetics Zebrafish* / genetics Zebrafish* / metabolism |
| IF | 4.096 |
| Resource | |
| Zebrafish | Tg(EF1a:ERK Biosensor-nes) (teen) |