Background: Immune checkpoint inhibitors have been widely used in the systematic treatment of various advanced solid tumors, but they show poor efficacy or lead to hyperprogression in some patients. Some studies reported that EGFR mutations, ALK fusion, interferon pathway or Wnt signal pathway gene mutations might reduce the efficacy of immunotherapy and lead to tumor hyperprogression. The aim of this study was to determine the frequency of gene variations associated with immunotherapy resistance or hyperprogression in advanced solid tumors. Methods: The tumor tissues or blood samples of 111 patients with advanced solid tumors were collected. The tumor tissues were sequenced by targeted next-generation sequencing (55 genes, Illumina novaseq 6000), MSI (PCR) and PD-L1 detection (IHC). Blood cfdna was extracted and sequenced by targeted next-generation sequencing (55 genes, Illumina novaseq 6000). Results: A total of 111 patients with advanced solid tumors were included in the study, including 34 patients with colorectal cancer, 27 patients with hepatobiliary tumors, 18 patients with gastric cancer, 15 patients with urothelial carcinoma, and 17 patients with other solid tumors. Among the 82 samples tested for MSI, there were 77 cases of MSS (93.9%), 1 case of MSI-L (1.2%), and 4 cases of MSI-H (4.9%). 44 samples were tested for PD-L1 expression. 33 (75%) had PD-L1 expression < 1%, and 11 (25%) had PD-L1 expression greater than 1% but less than 49%. Among the genes related to immunotherapy, the mutations of genes related to cell growth or cell cycle regulation (EGFR, ALK, CDKN2A and CCND1) accounted for 3 (2.7%), 5 (4.5%), 12 (10.8%) and 5 (4.5%), respectively. The mutation of antigen presentation related gene (B2M) accounted for 3 (2.7%). The mutations of interferon signaling pathway related genes (JAK1, JAK2, JAK3 and IFNGR1) accounted for 2 (1.8%), 5 (4.5%), 1 (0.9%) and 1 (0.9%), respectively. The mutations in Wnt signaling pathway related genes (APC, AXIN1, CTNNB1) accounted for 41 (36.9%), 5 (4.5%) and 7 (6.3%). The mutations of tumor suppressor genes (PTEN, STK11 and PTPN2) accounted for 9 (8.1%), 1 (0.9%) and 3 (2.7%). Mutations in other genes (APLNR, DNMT3a, MDM2) accounted for 1 (0.9%), 6 (5.4%) and 7 (6.3%). Conclusions: Our study shows that in patients with advanced solid tumors, there are multiple gene variants related to drug resistance or hyperprogression of tumor immune checkpoint inhibitor treatment, suggesting that it may be the reason for the difference in the efficacy of immunotherapy. It is necessary to further study the efficacy of immunotherapy in patients with different gene mutations and the combination of immune checkpoint inhibitors for patients with different pathway mutations.