Characterization of Two New Imatinib-Responsive Fusion Genes Generated by Disruption of PDGFRB in Eosinophilia-Associated Chronic Myeloproliferative Disorders.
Authors: Walz Christoph , Metzgeroth Georgia , Schoch Claudia , Haferlach Torsten , Hehlmann Rudiger , Hochhaus Andreas , Cross Nicholas C.P. , Reiter Andreas 
Published: April 2020

Fusion genes involving PDGFRA, PDGFRB, FGFR1 and JAK2 are seen in a substantial number of patients with BCR-ABL negative myeloproliferative disorders (MPD) and result in constitutive activation of the corresponding tyrosine kinase moiety. The vast majority of tyrosine kinase fusion partners contain coiled-coil domains or other dimerization motifs properties that are essential for malignant transformation. We have identified two patients presenting with eosinophilia-associated MPD and a t(5;12)(q31;q24) or a complex translocation t(1;5;11) with involvement of 5q31, respectively, suggesting a possible involvement of the PDGFRB gene which is located at chromosome band 5q31–33. 5′-rapid amplification of cDNA ends (5′-RACE) for the t(5;12) identified an in-frame mRNA fusion between ’G protein-coupled receptor kinase interactor 2′ (GIT2) exon 12 at 12q24 and PDGFRB exon 11. GIT2 is a member of the GIT protein family that is extensively alternative spliced in many distinct forms causing its functional diversity. A reciprocal transcript was amplified by RT-PCR with a fusion between PDGFRB exon 10 and GIT2 exon 13. GIT2-PDGFRB is predicted to be translated into a 742 amino acid fusion protein that retains the GIT2 N-terminal protein-protein interaction motif Ankyrin and an Arf GTPase activating protein (ArfGAP) domain fused to the transmembrane and catalytic domain of PDGFRB. The truncated GIT2 protein lacks coiled-coil domains as they are lost in the fusion protein due to the breakpoint within GIT2 intron 12. We therefore speculate that the Ankyrin repeat, which is one of the most common protein-protein interaction motifs in nature, may have replaced the function of a coiled-coil domain offering dimerization properties to the fusion protein. 5′-RACE for the complex t(1;5;11) identified an in-frame mRNA fusion between ’GPI-anchored membrane protein 1′ (GPIAP1) exon 7 at 11p13 and PDGFRB exon 11. Normal GPIAP1 is a cytoplasmic phosphoprotein which plays a mainly uncharacterized role in cellular activation or proliferation. The chimeric mRNA is predicted to encode an 803 amino acid fusion protein retaining the coiled-coil domain of GPIAP1 fused to the transmembrane and catalytic domains of PDGFRB. Both patients have been treated with 400 mg/day imatinib, which is a selective inhibitor of PDGFRB, and achieved rapid complete clinical and hematological remission. Residual GIT2-PDGFRB transcripts could be detected repeatedly during a 17 months follow up in case 1 whereas no follow-up samples have been available for case 2. These data give further evidence that numerous partner genes fuse to PDGFRB in BCR-ABL negative MPDs. In addition, the data demonstrate that cytogenetic analysis is a mandatory technique for the identification of tyrosine kinase fusion genes. In cases with abnormalities of chromosome 5q, a possible involvement of PDGFRB should be screened by adequate FISH and PCR-based techniques. Although their occurrence is rare in general, the identification of these fusion genes is essential for the successful treatment with tyrosine kinase inhibitors.