本帖最后由 StephenW 于 2017-2-1 14:46 编辑
An RNA-based signature enables high specificity detection of circulating tumor cells in hepatocellular carcinoma - Mark Kalinicha,1,
- Irun Bhana,b,1,
- Tanya T. Kwana,
- David T. Miyamotoa,c,
- Sarah Javaida,2,
- Joseph A. LiCausia,
- John D. Milnera,
- Xin Honga,
- Lipika Goyala,d,
- Srinjoy Sila,
- Melissa Choza,
- Uyen Hoa,
- Ravi Kapure,
- Alona Muzikanskya,f,
- Huidan Zhangg,
- David A. Weitzg,
- Lecia V. Sequista,d,
- David P. Ryana,d,
- Raymond T. Chungb,
- Andrew X. Zhua,d,
- Kurt J. Isselbachera,b,3,
- David T. Tinga,d,
- Mehmet Tonere,h,
- Shyamala Maheswarana,h,3, and
- Daniel A. Habera,d,i,3
- aMassachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129;
- bDivision of Gastroenterology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114;
- cDepartment of Radiation Oncology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114;
- dDivision of Hematology Oncology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114;
- eCenter for Bioengineering in Medicine, Massachusetts General Hospital and Harvard Medical School and Shriners Hospital for Children, Boston, MA 02114;
- fDivision of Biostatistics, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114;
- gDepartment of Physics, School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138;
- hDepartment of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114;
- iHoward Hughes Medical Institute, Chevy Chase, MD 20815
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Contributed by Kurt J. Isselbacher, December 11, 2016 (sent for review October 14, 2016; reviewed by Anil K. Rustgi and Timothy Wang)
Significance The early detection of hepatocellular carcinoma (HCC) is of paramount importance for improving patient outcomes, yet an accurate, high-throughput screening methodology has yet to be developed. By combining microfluidic depletion of hematopoietic cells from blood specimens with absolute quantification of lineage-derived transcripts, we demonstrate the highly specific detection of circulating tumor cells, enabling noninvasive detection and clinical monitoring of HCC.
Abstract Circulating tumor cells (CTCs) are shed into the bloodstream by invasive cancers, but the difficulty inherent in identifying these rare cells by microscopy has precluded their routine use in monitoring or screening for cancer. We recently described
a high-throughput microfluidic CTC-iChip, which efficiently depletes hematopoietic cells from blood specimens and enriches for CTCs with well-preserved RNA. Application of RNA-based digital PCR to detect CTC-derived signatures may thus enable highly accurate tissue lineage-based cancer detection in blood specimens. As proof of principle, we examined hepatocellular carcinoma (HCC), a cancer that is derived from liver cells bearing a unique gene expression profile. After identifying a digital signature of 10 liver-specific transcripts, we used a cross-validated logistic regression model to identify the presence of HCC-derived CTCs in nine of 16 (56%) untreated patients with HCC versus one of 31 (3%) patients with nonmalignant liver disease at risk for developing HCC (P < 0.0001). Positive CTC scores declined in treated patients: Nine of 32 (28%) patients receiving therapy and only one of 15 (7%) patients who had undergone curative-intent ablation, surgery, or liver transplantation were positive. RNA-based digital CTC scoring was not correlated with the standard HCC serum protein marker alpha fetoprotein (P = 0.57). Modeling the sequential use of these two orthogonal markers for liver cancer screening in patients with high-risk cirrhosis generates positive and negative predictive values of 80% and 86%, respectively. Thus, digital RNA quantitation constitutes a sensitive and specific CTC readout, enabling high-throughput clinical applications, such as noninvasive screening for HCC in populations where viral hepatitis and cirrhosis are prevalent.
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发表于 2017-2-1 14:43