title: An RNA-based signature enables high specificity detection of circulating tumor cells in hepatocellular carcinoma.
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.
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.
authors.forename: Mark
authors.initials: M
authors.lastname: Kalinich
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.|Division of Gastroenterology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114.
authors.forename: Irun
authors.initials: I
authors.lastname: Bhan
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.
authors.forename: Tanya T
authors.initials: TT
authors.lastname: Kwan
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.|Department of Radiation Oncology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114.
authors.forename: David T
authors.initials: DT
authors.lastname: Miyamoto
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.
authors.forename: Sarah
authors.initials: S
authors.lastname: Javaid
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.
authors.forename: Joseph A
authors.initials: JA
authors.lastname: LiCausi
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.
authors.forename: John D
authors.initials: JD
authors.lastname: Milner
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.
authors.forename: Xin
authors.initials: X
authors.lastname: Hong
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.|Division of Hematology Oncology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114.
authors.forename: Lipika
authors.initials: L
authors.lastname: Goyal
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.
authors.forename: Srinjoy
authors.initials: S
authors.lastname: Sil
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.
authors.forename: Melissa
authors.initials: M
authors.lastname: Choz
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.
authors.forename: Uyen
authors.initials: U
authors.lastname: Ho
authors.affiliation: Center for Bioengineering in Medicine, Massachusetts General Hospital and Harvard Medical School and Shriners Hospital for Children, Boston, MA 02114.
authors.forename: Ravi
authors.initials: R
authors.lastname: Kapur
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.|Division of Biostatistics, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114.
authors.forename: Alona
authors.initials: A
authors.lastname: Muzikansky
authors.affiliation: Department of Physics, School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138.
authors.forename: Huidan
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Physics, School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138.
authors.forename: David A
authors.initials: DA
authors.lastname: Weitz
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.|Division of Hematology Oncology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114.
authors.forename: Lecia V
authors.initials: LV
authors.lastname: Sequist
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.|Division of Hematology Oncology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114.
authors.forename: David P
authors.initials: DP
authors.lastname: Ryan
authors.affiliation: Division of Gastroenterology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114.
authors.forename: Raymond T
authors.initials: RT
authors.lastname: Chung
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.|Division of Hematology Oncology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114.
authors.forename: Andrew X
authors.initials: AX
authors.lastname: Zhu
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129; dhaber@mgh.harvard.edu maheswaran@helix.mgh.harvard.edu kisselbacher@mgh.harvard.edu.|Division of Gastroenterology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114.
authors.forename: Kurt J
authors.initials: KJ
authors.lastname: Isselbacher
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129.|Division of Hematology Oncology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114.
authors.forename: David T
authors.initials: DT
authors.lastname: Ting
authors.affiliation: Center for Bioengineering in Medicine, Massachusetts General Hospital and Harvard Medical School and Shriners Hospital for Children, Boston, MA 02114.|Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114.
authors.forename: Mehmet
authors.initials: M
authors.lastname: Toner
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129; dhaber@mgh.harvard.edu maheswaran@helix.mgh.harvard.edu kisselbacher@mgh.harvard.edu.|Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114.
authors.forename: Shyamala
authors.initials: S
authors.lastname: Maheswaran
authors.affiliation: Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129; dhaber@mgh.harvard.edu maheswaran@helix.mgh.harvard.edu kisselbacher@mgh.harvard.edu.|Division of Hematology Oncology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114.|Howard Hughes Medical Institute, Chevy Chase, MD 20815.
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Haber
mesh_terms: D014408:Biomarkers, Tumor / Q000097:blood*; D006528:Carcinoma, Hepatocellular / Q000097:blood / Q000175:diagnosis* / Q000473:pathology / Q000628:therapy; D019070:Cell Lineage; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D055088:Early Detection of Cancer / Q000379:methods*; D056945:Hep G2 Cells; D019694:Hepatitis B, Chronic / Q000097:blood; D059014:High-Throughput Nucleotide Sequencing / Q000295:instrumentation / Q000379:methods; D057166:High-Throughput Screening Assays; D006801:Humans; D056656:Lab-On-A-Chip Devices; D008103:Liver Cirrhosis / Q000097:blood; D008113:Liver Neoplasms / Q000097:blood / Q000175:diagnosis* / Q000473:pathology / Q000628:therapy; D016015:Logistic Models; D009360:Neoplastic Cells, Circulating; D011230:Precancerous Conditions / Q000097:blood; D011237:Predictive Value of Tests; D012333:RNA, Messenger / Q000097:blood*; D012334:RNA, Neoplasm / Q000097:blood*; D017423:Sequence Analysis, RNA / Q000295:instrumentation / Q000379:methods; D059010:Single-Cell Analysis; D059467:Transcriptome
chemical_list: D014408:Biomarkers, Tumor; D012333:RNA, Messenger; D012334:RNA, Neoplasm
keywords: blood biopsy; circulating tumor cells; early cancer detection; hepatocellular carcinoma; predictive modeling