OPENchip: an on-chip in situ molecular profiling platform for gene expression analysis and oncogenic mutation detection in single circulating tumour cells
| Autor: | Amos Chungwon Lee, Yunjin Jeong, Evangelia Darai, Seo Woo Song, Ahyoun Choi, Yeongjae Choi, Sumin Lee, Narayanan Madaboosi, Yongju Lee, Caleb S. Lee, Mats Nilsson, Hyung Jong Bae, Okju Kim, Wonshik Han, Sunghoon Kwon, Jessica Svedlund, Han-Byoel Lee, Huiran Yeom, Jin-Young Jang, Sung Min Kim, Daewon Lee, Dong Soon Lee |
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| Rok vydání: | 2020 |
| Předmět: |
In situ
Biomedical Engineering Gene Expression Bioengineering Breast Neoplasms Computational biology Biology Gene mutation medicine.disease_cause Biochemistry 03 medical and health sciences 0302 clinical medicine Gene expression medicine Humans Liquid biopsy Gene 030304 developmental biology Oligonucleotide Array Sequence Analysis 0303 health sciences Liquid Biopsy General Chemistry medicine.disease Neoplastic Cells Circulating Metastatic breast cancer 3. Good health Rolling circle replication 030220 oncology & carcinogenesis Mutation Female KRAS |
| Zdroj: | Lab on a chip. 20(5) |
| ISSN: | 1473-0189 |
| Popis: | Liquid biopsy holds promise towards practical implementation of personalized theranostics of cancer. In particular, circulating tumour cells (CTCs) can provide clinically actionable information that can be directly linked to prognosis or therapy decisions. In this study, gene expression patterns and genetic mutations in single CTCs are simultaneously analysed by strategically combining microfluidic technology and in situ molecular profiling technique. Towards this, the development and demonstration of the OPENchip (On-chip Post-processing ENabling chip) platform for single CTC analysis by epithelial CTC enrichment and subsequent in situ molecular profiling is reported. For in situ molecular profiling, padlock probes that identify specific desired targets to examine biomarkers of clinical relevance in cancer diagnostics were designed and used to create libraries of rolling circle amplification products. We characterize the OPENchip in terms of its capture efficiency and capture purity, and validate the probe design using different cell lines. By integrating the obtained results, molecular analyses of CTCs from metastatic breast cancer (HER2 (ERBB2) gene expression and PIK3CA mutations) and metastatic pancreatic cancer (KRAS gene mutations) patients were demonstrated without any off-chip processes. The results substantiate the potential implementation of early molecular detection of cancer through sequencing-free liquid biopsy. |
| Databáze: | OpenAIRE |
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