Abstract Title: ANGIOGENESIS SOLUBLE FACTORS AS HCC MARKERS FOR MONITORING HCV CIRRHOTIC PATIENTS
Valeria Mas, PhD1, Robert Fisher, MD1, Kellie Archer, PhD2, Kenneth Yanek, MS1 and Daniel Maluf, MD1. 1Surgery, Hume Lee Transplant Center, VCUHS, Richmond, VA, United States and 2Biostatistics, VCUHS, Richmond, VA, United States.
Body: Background. Angiogenesis (Ang) occurs in association with both healthy (liver regeneration) and pathologic (hepatocellular carcinoma (HCC)) conditions. Ang soluble factors are key mediators and may provide biologic insight in HCC. We aimed to identify different patterns of Ang gene expression in HCV tissues with and without HCC; and to evaluate the utility of Ang soluble factors as non-invasive markers of HCC.
Patients and Methods. Thirty-eight HCV-HCC tumors (2 T1N0M0, 17 T2N0M0, and 19 T3N0M0), 42 HCV cirrhotic and 6 normal liver tissues were studied using high-density oligonucleotide arrays. The robust-multiarray average method was used to estimate probe set (Pset) expression summaries. The significance analysis of microarrays method was used to identify Pset differentially expressed while controlling for the false discovery rate. Human Angiogenesis Microarray (HAM) was used for the protein detection of the following soluble angiogenic factors: EGF; TIMP-1; TIMP-2; HGF; Angiopn-1; Angiopn-2; VEGF-A; IP-10; PDGF; KGF; Angiogenin; VEGF-D; ICAM-1; FGF in plasma from 20 patients (Pts)(15 HCCs and 5 HCV cirrhotic Pts). This antibody array contains sixteen identical arrays of 14 capture antibodies in quadruplicate. Array detection is achieved with the addition of Streptavidin-Cy5 for fluorescent signals.
Results. From the analysis of the HCV-HCC vs. normal livers we found an important number of genes related to Ang differentially expressed including VGEF, PDGF, AGPTL2, ANG, EGFL6, EGFR, ANGPT1, ANGPT2, ICAM5, ICAM2, TIMP-2, among others (=0.01). Ang genes were also differentially expressed in HCV-HCC vs. HCV cirrhotic tissue analysis (VGEF, EGFL3, EGFR, VEGFB, among others) (=0.01). The protein detection assay in plasma samples using the HAM, showed that IP-10 (P=0.032), EGF (P=0.05), ICAM (P=0.01), KGF (P=0.052) were statistically significant different between HCV-HCC and HCV plasma samples. We also observed that the level of Angiopn-2 was related to the stage of the HCC tumor (P=0.02).
Conclusions. Differentially expressed Ang genes were observed between HCV Pts with and without HCC in liver tissue. Furthermore, we observed a relationship between angiogenic hepatic tumor tissue gene expression and plasma protein markers. The use of plasma levels of angiogenic factors as HCC biomarkers will be studied as a new tool for non-invasive diagnostic monitoring of HCC disease.