Familial colorectal cancer (CRC) is only partially explained by known germline predisposing genes. We performed whole-genome sequencing in 15 Polish families of many affected individuals, without mutations in known CRC predisposing genes. We focused on loss-of-function variants and functionally characterized them. We identified a frameshift variant in the CYBA gene (c.246delC) in one family and a splice site variant in the TRPM4 gene (c.25–1 G > T) in another family. While both variants were absent or extremely rare in gene variant databases, we identified four additional Polish familial CRC cases and two healthy elderly individuals with the CYBA variant (odds ratio 2.46, 95% confidence interval 0.48–12.69). Both variants led to a premature stop codon and to a truncated protein. Functional characterization of the variants showed that knockdown of CYBA or TRPM4 depressed generation of reactive oxygen species (ROS) in LS174T and HT-29 cell lines. Knockdown of TRPM4 resulted in decreased MUC2 protein production. CYBA encodes a component in the NADPH oxidase system which generates ROS and controls, e.g., bacterial colonization in the gut. Germline CYBA variants are associated with early onset inflammatory bowel disease, supported with experimental evidence on loss of intestinal mucus barrier function due to ROS deficiency. TRPM4 encodes a calcium-activated ion channel, which, in a human colonic cancer cell line, controls calcium-mediated secretion of MUC2, a major component of intestinal mucus barrier. We suggest that the gene defects in CYBA and TRPM4 mechanistically involve intestinal barrier integrity through ROS and mucus biology, which converges in chronic bowel inflammation.

This study aims to compare the effectiveness and safety of anti-programmed cell death protein 1 (PD-1) immunotherapy and tyrosine kinase inhibitors (TKIs) combined with intra-arterial therapy (IAT) or systemic chemotherapy, and anti-PD-1 immunotherapy combined with IAT or systemic chemotherapy in patients with advanced biliary tract carcinoma (BTC). Patients with BTC who received IAT combined with anti-PD-1 immunotherapy and/or TKIs or the combination of systemic chemotherapy with anti-PD-1 immunotherapy and/or TKIs were divided into 4 groups according to inclusion and exclusion criteria: the combination of IAT with TKIs and anti-PD-1 immunotherapy group (ITP group), the systemic chemotherapy combined with TKIs and anti-PD-1 immunotherapy group (CTP group), the combination of IAT with anti-PD-1 immunotherapy group (IP group) and the systemic chemotherapy combined with anti-PD-1 immunotherapy group (CP group). The baseline characteristics, progression-free survival (PFS), treatment efficacy and adverse reactions were compared between ITP and CTP groups and between IP and CP groups, and risk factors related to PFS were analyzed. The median PFS in ITP group was 3.975 months, and the 6-month-PFS was 31.6%; the median PFS in CTP group was 3.285 months, and the 6-month-PFS was 11.1%. There was a statistical difference between ITP group and CTP group. The median PFS in the IP and CP groups were 4.534 months and 2.267 months, respectively, which were also statistically significant. The objective response rate and disease control rate (DCR) of ITP and CTP groups were not statistically significant. There was no significant difference in objective response rate and DCR between IP group and CP group. The lymph node metastasis was an independent risk factor for PFS. Compared with the CTP group, the ITP group had a higher incidence of post-treatment fever and difficulty urination, and the IP group had a higher incidence of 3 degree AST elevation and difficulty urination than the CP group. IAT combined with anti-PD-1 immunotherapy and TKIs or IAT combined with anti-PD-1 immunotherapy for advanced BTC were relatively effective and safe. The lymph node metastasis was an independent risk factor affecting the prognosis of advanced BTC.

Familial colorectal cancer (CRC) is only partially explained by known germline predisposing genes. We performed whole genome sequencing in 15 Polish families of many affected individuals, without mutations in known CRC predisposing genes. We focused on loss-of-function variants and functionally characterized them. We identified a frameshift variant in the CYBA gene (c.246delC) in one family and a splice site variant in the TRPM4 gene (c.25-1 G>T) in another family. While both variants were absent or extremely rare in gene variant databases, we identified four additional Polish familial CRC cases and two healthy elderly individuals with the CYBA variant (odds ratio 2.46, 95% confidence interval 0.48-12.69). Both variants led to a premature stop codon and to a truncated protein. Functional characterization of the variants showed that knockdown of CYBA or TRPM4 depressed generation of reactive oxygen species (ROS) in LS174T and HT-29 cell lines. Knockdown of TRPM4 resulted in decreased MUC2 protein production. CYBA encodes a component in the NADPH oxidase system which generates ROS and controls, e.g., bacterial colonization in the gut. Germline CYBA variants are associated with early onset inflammatory bowel disease, supported with experimental evidence on loss of intestinal mucus barrier function due to ROS deficiency. TRPM4 encodes a calcium-activated ion channel, which in a human colonic cancer cell line controls calcium-mediated secretion of MUC2, a major component of intestinal mucus barrier. We suggest that the gene defects in CYBA and TRPM4 mechanistically involve intestinal barrier integrity through ROS and mucus biology, which converges in chronic bowel inflammation.