Autophagosome formation is a fundamental process in macroautophagy/autophagy, a conserved self-eating mechanism in all eukaryotes, which requires the conjugating ATG (autophagy related) protein complex, ATG12–ATG5-ATG16L1 and lipidated MAP1LC3/LC3 (microtubule associated protein 1 light chain 3). How the ATG12–ATG5-ATG16L1 complex is recruited to membranes is not fully understood. Here, we demonstrated that RAB33B plays a key role in recruiting the ATG16L1 complex to phagophores during starvation-induced autophagy. Crystal structures of RAB33B bound to the coiled-coil domain (CCD) of ATG16L1 revealed the recognition mechanism between RAB33B and ATG16L1. ATG16L1 is a novel RAB-binding protein (RBP) that can induce RAB proteins to adopt active conformation without nucleotide exchange. RAB33B and ATG16L1 mutually determined the localization of each other on phagophores. RAB33B-ATG16L1 interaction was required for LC3 lipidation and autophagosome formation. Upon starvation, a fraction of RAB33B translocated from the Golgi to phagophores and recruited the ATG16L1 complex. In this work, we reported a new mechanism for the recruitment of the ATG12–ATG5-ATG16L1 complex to phagophores by RAB33B, which is required for autophagosome formation. Abbreviations : ATG: autophagy-related; Cα: alpha carbon; CCD: coiled-coil domain; CLEM: correlative light and electron microscopy; DTE: dithioerythritol; EBSS: Earle’s balanced salt solution; EDTA: ethylenediaminetetraacetic acid; EGFP: enhanced green fluorescent protein; FBS: fetal bovine serum; FLIM: fluorescence lifetime imaging microscopy; FRET: Förster resonance energy transfer; GDP: guanosine diphosphate; GOLGA2/GM130: golgin A2; GppNHp: guanosine 5ʹ-[β,γ-imido]triphosphate; GST: glutathione S-transferase; GTP: guanosine triphosphate; GTPγS: guanosine 5ʹ-O-[gamma-thio]triphosphate; HA (tag): hemagglutinin (tag); HEK: human embryonic kidney; HeLa: Henrietta Lacks; HEPES: (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid); IgG: immunoglobulin G; K_d: dissociation constant; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MCF7: Michigan cancer foundation-7; MEF: mouse embryonic fibroblast; MEM: minimum essential medium Eagle; MST: microscale thermophoresis; NEAA: non-essential amino acids; PBS: phosphate-buffered saline; PE: phosphatidylethanolamine; PtdIns3P: phosphatidylinositol-3-phosphate; RAB: RAS-associated binding; RB1CC1/FIP200: RB1 inducible coiled-coil protein 1; RBP: RAB-binding protein; SD: standard deviation; SDS: sodium dodecyl sulfate; SQSTM1/p62: sequestosome 1; TBS-T: tris-buffered saline-tween 20; WD (repeat): tryptophan-aspartic acid (repeat); WIPI2B: WD repeat domain phosphoinositide interacting 2B; WT: wild type

A postoperative scoring system for post-hepatectomy early recurrence of colorectal liver metastases A postoperative scoring system for post-hepatectomy early recurrence of colorectal liver metastases

Objective: To investigate the impact of postoperative infectious complications (POI) on the long-term outcomes of patients with colorectal cancer liver metastasis (CRLM) after simultaneous resection of colorectal cancer and liver metastases.Methods: Four hundred seventy-nine CRLM patients receiving simultaneous resection between February 2010 and February 2018 at our hospital were enrolled. A 1:3 propensity score matching analysis (PSM) analysis was performed to balance covariates and avoid selection bias. After PSM, 90 patients were distributed to the POI group, and 233 patients were distributed to the no POI group. A log-rank test was performed to compare the progression-free survival (PFS) and overall survival (OS) data. A multivariate Cox regression model was employed to identify prognostic factors influencing OS and PFS. A value of two-sided P<0.05 was considered statistically significant.Results: Compared to patients in the no POI group, patients in the POI group were more likely to have hepatic portal occlusion (78.9% vs. 66.3%, P=0.021), operation time ≥325 min (61.1% vs. 48.1%, P=0.026), and intraoperative blood loss ≥200 ml (81.1% vs. 67.6%, P=0.012). In multivariate analysis, intraoperative blood loss ≥200 ml (OR = 2.057, 95% CI: 1.165-3.634, P=0.013) was identified as the only independent risk factor for POI. Patients with POI had a worse PFS (P&lt;0.001, median PFS: 7.5 vs. 12.7 months) and a worse OS (P=0.010, median OS: 38.8 vs. 59.0 months) than those without POI. After 1:3 PSM analysis, no differences in clinicopathologic parameters were detected between the POI group and the no POI group. Patients with POI had a worse PFS (P=0.013, median PFS: 7.5 vs. 11.1 months) and a worse OS (P=0.020, median OS: 38.8 vs. 59.0 months) than those without POI. Multivariate analysis showed that POI was an independent predictor for worse PFS (HR=1.410, 95% CI: 1.065-1.869, P=0.017) and worse OS (HR=1.682, 95% CI: 1.113-2.544, P=0.014).Conclusions: POI can significantly worsen the long-term outcomes of CRLM patients receiving simultaneous resection of colorectal cancer and liver metastases and should be considered to improve postoperative management and make better treatment decisions for these patients.

Background: Intrahepatic cholangiocarcinoma (ICC) is a highly aggressive primary liver malignancy, with poor long-term outcomes even after curative-intent resection. Postoperative adjuvant chemotherapy (pAC) is increasingly used, but its benefit is not uniform across all patients. The systemic immune-inflammation index (SII) has emerged as a potential prognostic marker in several cancers, but its role in ICC remains unclear. Methods: We retrospectively analyzed 445 ICC patients who underwent R0 hepatic resection at a single tertiary center between 2000 and 2023. Preoperative SII was calculated, and patients were stratified into high- and low-SII groups. The impact of SII on overall survival (OS) and recurrence-free survival (RFS) was evaluated, along with its interaction with pAC. Multivariate Cox regression models and maximally selected rank statistics were used for analysis. Results: The median follow-up was 34.3 months. High SII independently predicted worse OS and RFS (p < 0.001), outperforming conventional inflammatory and nodal indices. Lymph node ratio (LNR) also independently predicted survival but did not modify the effect of pAC. Interaction analysis revealed that pAC significantly improved OS in high-SII patients (5-year OS: 33% with pAC vs. 23% without; HR 0.62, 95% CI 0.42–0.94, p = 0.022) but conferred no significant benefit in low-SII patients (5-year OS: 49% with pAC vs. 55% without; HR 0.71, 95% CI 0.48–1.05, p = 0.089). Conclusions: SII is a robust prognostic biomarker in ICC and can guide individualized postoperative therapy. High-SII patients derive substantial survival benefit from adjuvant chemotherapy, whereas low-SII patients may be spared unnecessary treatment. Integrating SII into postoperative risk stratification may optimize outcomes and reduce overtreatment in ICC.