PMC7568639_01
Timeline of clinical treatment course and radiologic features. (A) Timeline of the patient's clinical treatment course. (B-I) Dynamic imaging of lung lesions and bone metastases at different stages of crizotinib treatment. Computed tomographic (CT) images of the tumor in the left upper lobe (B) and the enlarged mediastinal lymph nodes (E) prior to crizotinib treatment. Primary tumor (C and D) and mediastinal lymph nodes (F and G) eliminated in lung after 2 months and 9 months of crizotinib treatment, respectively. Bone scintigraphy images of the patient's multiple osteoblastic metastasis before (H) and 9 months after crizotinib treatment (I). Red arrows indicate the tumor and lymph node sites.
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A 45-year-old Chinese female was admitted to the hospital in March 2019 with a 2-month history of cough and white sticky phlegm (Figure 1A). The chest computed tomography (CT) scan showed an irregular mass in the left upper lung lobe with multiple mediastinal lymph nodes enlargement (Figure 1B and E). 99mTc-Methylene Diphosphate (MDP) bone scintigraphy images revealed that osseous destruction in the patient was characterized by multiple osteoblastic metastases, including the head of the right humerus, multiple ribs, multiple vertebras, pelvis and femurs (Figure 1H). Two months later (June 2019), a complete response in the primary lung tumor was achieved according to chest CT scan imaging, showing that the left upper lobe mass and metastatic lymph nodes were eliminated (Figure 1C and F). After 9 months of crizotinib treatment (January 2020), no tumor and no metastasis were found in lung (Figure 1D and G), and bone scintigraphy images demonstrated that multiple bone metastatic lesions were significantly relieved (Figure 1I).
Identification and verification of a novel MPRIP-ROS1 fusion. (A) RNA sequencing reads indicating MPRIP and ROS1 fusion regions were visualized by the Integrative Genomic Viewer (IGV) software. The fusion breakpoints are localized at chr17: p11.2: 17,080,726 and chr6: q22.1: 117,642,557, respectively. (B) Schematic of genomic rearrangement involving the fusion breakpoints in mRNA and protein level. The transcript resulted in exons 1-21 of MPRIP containing coiled-coil domains fused to exons 35-43 of ROS1 including kinase domain. Pink, MPRIP; Blue, ROS1; CC, coiled-coil domains; KD, kinase domain; e, exon. (C) Amplification products of fusion region by reverse transcriptase polymerase chain reaction (RT-PCR) using two forward primers located in exon 21 of MPRIP and two reverse primers located in exon 35 of ROS1. (D) Sanger sequencing chromatogram of the RT-PCR products further confirms the breakpoints and fusion sequence.
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A novel ROS1 rearrangement involving myosin phosphatase Rho interacting protein (MPRIP) gene was identified by RNA fusion panel based on amplicon sequencing (Berry Oncology Corporation), which revealed the harboring of MPRIP exon 21-ROS1 exon 35 rearrangement (Figure 2A and B). The breakpoints and fusion sequence were further confirmed using reverse transcriptase polymerase chain reaction (RT-PCR) followed by Sanger sequencing (Figure 2C and D).