Gene Expression in Fixed Tissues and Outcome in Hepatocellular Carcinoma

Yujin Hoshida, M.D., Ph.D., Augusto Villanueva, M.D., Masahiro Kobayashi, M.D., Judit Peix, A.S., Derek Y. Chiang, Ph.D., Amy Camargo, B.A., Supriya Gupta, B.S., Jamie Moore, M.A., B.S., Matthew J. Wrobel, M.S., Jim Lerner, B.S., Michael Reich, B.S., Jennifer A. Chan, M.D., Jonathan N. Glickman, M.D., Ph.D., Kenji Ikeda, M.D., Masaji Hashimoto, M.D., Goro Watanabe, M.D., Maria G. Daidone, Ph.D., Sasan Roayaie, M.D., Myron Schwartz, M.D., Swan Thung, M.D., Helga B. Salvesen, M.D., Ph.D., Stacey Gabriel, Ph.D., Vincenzo Mazzaferro, M.D., Jordi Bruix, M.D., Scott L. Friedman, M.D., Hiromitsu Kumada, M.D., Josep M. Llovet, M.D., and Todd R. Golub, M.D.

ABSTRACT

Background It is a challenge to identify patients who, after undergoing potentially curative treatment for hepatocellular carcinoma, are at greatest risk for recurrence. Such high-risk patients could receive novel interventional measures. An obstacle to the development of genome-based predictors of outcome in patients with hepatocellular carcinoma has been the lack of a means to carry out genomewide expression profiling of fixed, as opposed to frozen, tissue.

Methods We aimed to demonstrate the feasibility of gene-expression profiling of more than 6000 human genes in formalin-fixed, paraffin-embedded tissues. We applied the method to tissues from 307 patients with hepatocellular carcinoma, from four series of patients, to discover and validate a gene-expression signature associated with survival.

Results The expression-profiling method for formalin-fixed, paraffin-embedded tissue was highly effective: samples from 90% of the patients yielded data of high quality, including samples that had been archived for more than 24 years. Gene-expression profiles of tumor tissue failed to yield a significant association with survival. In contrast, profiles of the surrounding nontumoral liver tissue were highly correlated with survival in a training set of tissue samples from 82 Japanese patients, and the signature was validated in tissues from an independent group of 225 patients from the United States and Europe (P=0.04).

Conclusions We have demonstrated the feasibility of genomewide expression profiling of formalin-fixed, paraffin-embedded tissues and have shown that a reproducible gene-expression signature correlated with survival is present in liver tissue adjacent to the tumor in patients with hepatocellular carcinoma.

Peginterferon Alfa-2a and Ribavirin in Latino and Non-Latino Whites with Hepatitis C

Maribel Rodriguez-Torres, M.D., Lennox J. Jeffers, M.D., Muhammad Y. Sheikh, M.D., Lorenzo Rossaro, M.D., Victor Ankoma-Sey, M.D., Fayez M. Hamzeh, M.D., Ph.D., Paul Martin, M.D., for the Latino Study Group

ABSTRACT

Background Race has been shown to be a factor in the response to therapy for hepatitis C virus (HCV) infection, and limited data suggest that ethnic group may be as well; however, Latinos and other ethnic subpopulations have been underrepresented in clinical trials. We evaluated the effect of Latino ethnic background on the response to treatment with peginterferon alfa-2a and ribavirin in patients infected with HCV genotype 1 who had not been treated previously.

Methods In a multicenter, open-label, nonrandomized, prospective study, 269 Latino and 300 non-Latino whites with HCV infection received peginterferon alfa-2a, at a dose of 180 µg per week, and ribavirin, at a dose of 1000 or 1200 mg per day, for 48 weeks, and were followed through 72 weeks. The primary end point was a sustained virologic response. We enrolled Latinos whose parents and grandparents spoke Spanish as their primary language; nonwhite Latinos were excluded.

Results Baseline characteristics were similar in the Latino and non-Latino groups, although higher proportions of Latino patients were 40 years of age or younger, had a body-mass index (BMI, the weight in kilograms divided by the square of the height in meters) of more than 27 or more than 30, and had cirrhosis. The rate of sustained virologic response was higher among non-Latino whites than among Latinos (49% vs. 34%, P<0.001).> of HCV RNA in serum was more frequent in non-Latino whites at week 4 (P=0.045) and throughout the treatment period (P<0.001 for all other comparisons). Latino or non-Latino background was an independent predictor of the rate of sustained virologic response in an analysis adjusted for baseline differences in BMI, cirrhosis, and other characteristics. Adherence to treatment did not differ significantly between the two groups. The numbers of patients with adverse events and dose modifications were similar in the two groups, but fewer Latino patients discontinued therapy because of adverse events.

Conclusions Treatment with peginterferon alfa-2a and ribavirin for 48 weeks resulted in rates of sustained virologic response among patients infected with HCV genotype 1 that were lower among Latino whites than among non-Latino whites. Strategies to improve the sustained virologic response in Latinos are needed. (ClinicalTrials.gov number, NCT00107653 [ClinicalTrials.gov] .)

Tenofovir Disoproxil Fumarate versus Adefovir Dipivoxil for Chronic Hepatitis B

Patrick Marcellin, M.D., E. Jenny Heathcote, M.D., Maria Buti, M.D., Ed Gane, M.D., Robert A. de Man, M.D., Zahary Krastev, M.D., George Germanidis, M.D., Sam S. Lee, M.D., Robert Flisiak, M.D., Kelly Kaita, M.D., Michael Manns, M.D., Iskren Kotzev, M.D., Konstantin Tchernev, M.D., Peter Buggisch, M.D., Frank Weilert, M.D., Oya Ovung Kurdas, M.D., Mitchell L. Shiffman, M.D., Huy Trinh, M.D., Mary Kay Washington, M.D., Jeff Sorbel, M.S., Jane Anderson, Ph.D., Andrea Snow-Lampart, B.S., Elsa Mondou, M.D., Joe Quinn, M.P.H., and Franck Rousseau, M.D.

ABSTRACT

Background Tenofovir disoproxil fumarate (DF) is a nucleotide analogue and a potent inhibitor of human immunodeficiency virus type 1 reverse transcriptase and hepatitis B virus (HBV) polymerase.

Methods In two double-blind, phase 3 studies, we randomly assigned patients with hepatitis B e antigen (HBeAg)–negative or HBeAg-positive chronic HBV infection to receive tenofovir DF or adefovir dipivoxil (ratio, 2:1) once daily for 48 weeks. The primary efficacy end point was a plasma HBV DNA level of less than 400 copies per milliliter (69 IU per milliliter) and histologic improvement (i.e., a reduction in the Knodell necroinflammation score of 2 or more points without worsening fibrosis) at week 48. Secondary end points included viral suppression (i.e., an HBV DNA level of <400> improvement, serologic response, normalization of alanine aminotransferase levels, and development of resistance mutations.

Results At week 48, in both studies, a significantly higher proportion of patients receiving tenofovir DF than of those receiving adefovir dipivoxil had reached the primary end point (P<0.001).> patients receiving tenofovir DF than patients receiving adefovir dipivoxil (93% vs. 63%, P<0.001)> patients receiving tenofovir DF than patients receiving adefovir dipivoxil (76% vs. 13%, P<0.001).> patients treated with tenofovir DF than those treated with adefovir dipivoxil had normalized alanine aminotransferase levels (68% vs. 54%, P=0.03) and loss of hepatitis B surface antigen (3% vs. 0%, P=0.02). At week 48, amino acid substitutions within HBV DNA polymerase associated with phenotypic resistance to tenofovir DF or other drugs to treat HBV infection had not developed in any of the patients. Tenofovir DF produced a similar HBV DNA response in patients who had previously received lamivudine and in those who had not. The safety profile was similar for the two treatments in both studies.

Conclusions Among patients with chronic HBV infection, tenofovir DF at a daily dose of 300 mg had superior antiviral efficacy with a similar safety profile as compared with adefovir dipivoxil at a daily dose of 10 mg through week 48. (ClinicalTrials.gov numbers, NCT00116805 [ClinicalTrials.gov] and NCT00117676 [ClinicalTrials.gov] .)