Hiroki Murai1, Takahiro Kodama1, Kazuki Maesaka1, Shoichiro Tange2, Daisuke Motooka3, Yutaka Suzuki4, Yasuyuki Shigematsu5, Kentaro Inamura5, Yoshihiro Mise6, Akio Saiura6, Yoshihiro Ono7, Yu Takahashi7, Yota Kawasaki8, Satoshi Iino9, Shogo Kobayashi10, Masashi Idogawa2, Takashi Tokino2, Tomomi Hashidate-Yoshida11, Hideo Shindou11,12, Masanori Miyazaki13, Yasuharu Imai14, Satoshi Tanaka15, Eiji Mita15, Kazuyoshi Ohkawa16, Hayato Hikita1, Ryotaro Sakamori1, Tomohide Tatsumi1, Hidetoshi Eguchi10, Eiichi Morii17, Tetsuo Takehara1. 1. Department of Gastroenterology and Hepatology, Osaka University Graduate School of Medicine, Suita, Japan. 2. Department of Medical Genome Sciences, Research Institute for Frontier Medicine, Sapporo Medical University School of Medicine, Sapporo, Japan. 3. Genome Information Research Center, Research Institute for Microbial Diseases, Osaka University, Suita, Japan. 4. Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Chiba, Japan. 5. Division of Pathology, Cancer Institute, Department of Pathology, Cancer Institute Hospital, Japanese Foundation for Cancer Research, Tokyo, Japan. 6. Department of Hepatobiliary-Pancreatic Surgery, Juntendo University School of Medicine, Tokyo, Japan. 7. Division of Hepatobiliary and Pancreatic Surgery, Cancer Institute Hospital, Japanese Foundation for Cancer Research, Tokyo, Japan. 8. Department of Digestive Surgery, Breast, and Thyroid Surgery, Graduate School of Medical Sciences, Kagoshima University, Kagoshima, Japan. 9. Department of Digestive Surgery, Kagoshima Principal Hospital, Kagoshima, Japan. 10. Department of Gastroenterological Surgery, Osaka University Graduate School of Medicine, Suita, Japan. 11. Department of Lipid Signaling, National Center for Global Health and Medicine, Tokyo, Japan. 12. Department of Lipid Medical Science, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan. 13. Department of Gastroenterology and Hepatology, Osaka Police Hospital, Osaka, Japan. 14. Department of Gastroenterology and Hepatology, Ikeda Municipal Hospital, Osaka, Japan. 15. Department of Gastroenterology and Hepatology, National Hospital Organization Osaka National Hospital, Osaka, Japan. 16. Department of Hepatobiliary and Pancreatic Oncology, Osaka International Cancer Institute, Osaka, Japan. 17. Department of Pathology, Osaka University Graduate School of Medicine, Osaka, Japan.
Abstract
BACKGROUND AND AIMS: Immunotherapy has become the standard-of-care treatment for hepatocellular carcinoma (HCC), but its efficacy remains limited. To identify immunotherapy-susceptible HCC, we profiled the molecular abnormalities and tumor immune microenvironment (TIME) of rapidly increasing nonviral HCC. APPROACHES AND RESULTS: We performed RNA-seq of tumor tissues in 113 patients with nonviral HCC and cancer genome sequencing of 69 genes with recurrent genetic alterations reported in HCC. Unsupervised hierarchical clustering classified nonviral HCCs into three molecular classes (Class I, II, III), which stratified patient prognosis. Class I, with the poorest prognosis, was associated with TP53 mutations, whereas class III, with the best prognosis, was associated with cadherin-associated protein beta 1 (CTNNB1) mutations. Thirty-eight percent of nonviral HCC was defined as an immune class characterized by a high frequency of intratumoral steatosis and a low frequency of CTNNB1 mutations. Steatotic HCC, which accounts for 23% of nonviral HCC cases, presented an immune-enriched but immune-exhausted TIME characterized by T cell exhaustion, M2 macrophage and cancer-associated fibroblast (CAF) infiltration, high PD-L1 expression, and TGF-β signaling activation. Spatial transcriptome analysis suggested that M2 macrophages and CAFs may be in close proximity to exhausted CD8+ T cells in steatotic HCC. An in vitro study showed that palmitic acid-induced lipid accumulation in HCC cells upregulated PD-L1 expression and promoted immunosuppressive phenotypes of cocultured macrophages and fibroblasts. Patients with steatotic HCC, confirmed by chemical-shift MR imaging, had significantly longer PFS with combined immunotherapy using anti-PD-L1 and anti-VEGF antibodies. CONCLUSIONS: Multiomics stratified nonviral HCCs according to prognosis or TIME. We identified the link between intratumoral steatosis and immune-exhausted immunotherapy-susceptible TIME.
BACKGROUND AND AIMS: Immunotherapy has become the standard-of-care treatment for hepatocellular carcinoma (HCC), but its efficacy remains limited. To identify immunotherapy-susceptible HCC, we profiled the molecular abnormalities and tumor immune microenvironment (TIME) of rapidly increasing nonviral HCC. APPROACHES AND RESULTS: We performed RNA-seq of tumor tissues in 113 patients with nonviral HCC and cancer genome sequencing of 69 genes with recurrent genetic alterations reported in HCC. Unsupervised hierarchical clustering classified nonviral HCCs into three molecular classes (Class I, II, III), which stratified patient prognosis. Class I, with the poorest prognosis, was associated with TP53 mutations, whereas class III, with the best prognosis, was associated with cadherin-associated protein beta 1 (CTNNB1) mutations. Thirty-eight percent of nonviral HCC was defined as an immune class characterized by a high frequency of intratumoral steatosis and a low frequency of CTNNB1 mutations. Steatotic HCC, which accounts for 23% of nonviral HCC cases, presented an immune-enriched but immune-exhausted TIME characterized by T cell exhaustion, M2 macrophage and cancer-associated fibroblast (CAF) infiltration, high PD-L1 expression, and TGF-β signaling activation. Spatial transcriptome analysis suggested that M2 macrophages and CAFs may be in close proximity to exhausted CD8+ T cells in steatotic HCC. An in vitro study showed that palmitic acid-induced lipid accumulation in HCC cells upregulated PD-L1 expression and promoted immunosuppressive phenotypes of cocultured macrophages and fibroblasts. Patients with steatotic HCC, confirmed by chemical-shift MR imaging, had significantly longer PFS with combined immunotherapy using anti-PD-L1 and anti-VEGF antibodies. CONCLUSIONS: Multiomics stratified nonviral HCCs according to prognosis or TIME. We identified the link between intratumoral steatosis and immune-exhausted immunotherapy-susceptible TIME.