Development of a novel ALK rearrangement screening test for non-small cell lung cancers
| Autor: | Shu Ching Yang, Nan Haw Chow, Hung Wen Tsai, Yi Lin Chen, Wan Li Chen, Yi Chia Cheng, Ming Ching Lin, Chung Liang Ho, Wu Chou Su, Chien Chung Lin |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Male
Genetic Screens Lung Neoplasms Tissue Fixation Pulmonology Gene Identification and Analysis Artificial Gene Amplification and Extension Cell Cycle Proteins Polymerase Chain Reaction Lung and Intrathoracic Tumors Fusion gene Cohort Studies Carcinoma Non-Small-Cell Lung hemic and lymphatic diseases Fusion Genes Medicine and Health Sciences Anaplastic lymphoma kinase Anaplastic Lymphoma Kinase Early Detection of Cancer Aged 80 and over Gene Rearrangement Multidisciplinary Paraffin Embedding medicine.diagnostic_test Fluorescent in Situ Hybridization Serine Endopeptidases ALK Gene Amplification DNA Neoplasm Middle Aged Reverse transcription polymerase chain reaction ErbB Receptors Oncology Immunohistochemistry Medicine Female Microtubule-Associated Proteins medicine.drug Research Article Adult medicine.drug_class Science Molecular Probe Techniques Biology Research and Analysis Methods Crizotinib Gene Types Formaldehyde medicine Genetics Humans Molecular Biology Techniques Molecular Biology Aged Cell Proliferation Base Sequence Biology and Life Sciences Cancers and Neoplasms Reverse Transcriptase-Polymerase Chain Reaction Probe Hybridization Non-Small Cell Lung Cancer Pleural Effusion Malignant ALK inhibitor Pleural Effusion HEK293 Cells Cancer research Cytogenetic Techniques Fluorescence in situ hybridization |
| Zdroj: | PLoS ONE, Vol 16, Iss 9, p e0257152 (2021) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | Approximately 5–7% of non–small cell lung cancer (NSCLC) cases harbor an anaplastic lymphoma kinase (ALK) fusion gene and may benefit from ALK inhibitor therapy. To detect ALK fusion genes, we developed a novel test using reverse transcription polymerase chain reaction (RT-PCR) for the ALK kinase domain (KD). Since ALK expression is mostly silenced in the adult with the exception of neuronal tissue, the normal lung tissue, mesothelial lining, and inflammatory cells are devoid of ALK transcript, making ALK KD RT-PCR an ideal surrogate test for ALK fusion transcripts in lung or pleural effusion. The test was designed with a short PCR product (197 bp) to work for both malignant pleural effusion (MPE) and formalin-fixed, paraffin-embedded (FFPE) NSCLC samples. Using ALK IHC as a reference, the sensitivity of the test was 100% for both MPE and FFPE. The specificity was 97.6% for MPE and 97.4% for FFPE. Two false positive cases were found. One was a metastatic brain lesion which should be avoided in the future due to intrinsic ALK expression in the neuronal tissue. The other one resulted from ALK gene amplification. Due to potential false positivity, subsequent confirmation tests such as fluorescence in situ hybridization or multiplex PCR would be preferable. Nevertheless, the test is simple and inexpensive with no false negativity, making it a desirable screening test. It also offers an advantage over multiplex RT-PCR with the capability to detect novel ALK fusions. Indeed through the screening test, we found a novel ALK fusion partner (sperm antigen with calponin homology and coiled-coil domains 1 like gene, SPECC1L) with increased sensitivity to crizotinib in vitro. In summary, a novel RNA-based ALK KD analysis was developed for ALK rearrangement screening in MPE and FFPE specimens of NSCLC. This simple inexpensive test can be implemented as routine diagnostics. |
| Databáze: | OpenAIRE |
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