Kirsten Remmert

Steatotic liver disease is common, yet the mechanisms by which hepatocytes cope with surges in dietary fatty acids remain unclear. Here we use single-cell tissue imaging (scPhenomics) and spatial proteomics to map lipid handling across dietary states. Fasting remodeled mitochondria and lipid droplets (LDs), increasing mitochondria–LD contacts, whereas contacts were infrequent in Western diet (WD)-fed male mice. Fasting also elevated perilipin-5 (PLIN5), a mediator of mitochondria-LD tethering. PLIN5 overexpression modulated contact formation in a phosphorylation-dependent manner: the S155A variant enhanced organelle contacts and LD expansion, whereas the S155E variant reduced contacts and yielded fewer, smaller LDs. Overexpression of the S155A variant in WD reduced lipotoxicity. These data reveal an adaptive organelle-interaction program that channels lipids during nutrient stress and is attenuated by an obesogenic diet. Our work establishes scPhenomics for spatially resolved cell-state analysis and identifies PLIN5 phosphorylation as a lever to tune hepatocyte lipid flux, suggesting therapeutic potential for targeting mitochondria–LD coupling.

Understanding protein distribution patterns across tissue architecture is crucial for deciphering organ function in health and disease. Here we show the application of single-cell Deep Visual Proteomics to perform spatially resolved proteome analysis of individual cells in native liver tissue. We built a robust framework comprising strategic cell selection and continuous protein gradient mapping, allowing the investigation of larger clinical cohorts. We generated a comprehensive spatial map of the human hepatic proteome by analysing hundreds of isolated hepatocytes from 18 individuals. Among the 2,500 proteins identified per cell, about half exhibited zonated expression patterns. Cross-species comparison with male mice revealed conserved metabolic functions and human-specific features of liver zonation. Analysis of samples with disrupted liver architecture demonstrated widespread loss of protein zonation, with pericentral proteins being particularly susceptible. Our study provides a comprehensive and open-access resource of human liver organization while establishing a broadly applicable framework for spatial proteomics analyses along tissue gradients.

PURPOSE: Most patients with metastatic colorectal cancer (CRC) have microsatellite stable or mismatch repair-proficient tumors, which are resistant to immunotherapy especially in cases of liver metastases. We sought to determine whether tumor-targeted interleukin-12 (PDS01ADC) can improve outcomes for patients with CRC liver metastases managed with hepatic artery infusion pump (HAIP) chemotherapy. MATERIALS AND METHODS: NCT05286814 is a phase II nonrandomized trial evaluating subcutaneous PDS01ADC in combination with HAIP floxuridine and systemic chemotherapy (leucovorin, 5-fluorouracil, and oxaliplatin or leucovorin, 5-fluorouracil, and irinotecan) in patients with unresectable microsatellite stable or mismatch repair-proficient colorectal liver metastases previously treated with at least one line of systemic chemotherapy. The primary end points for the planned interim analysis were overall response rate and safety. RESULTS: Nine patients were included in this planned interim analysis. Seventy-eight percent (7/9) of patients receiving PDS01ADC with HAIP therapy had partial or complete responses at 6 months. The median hepatic progression-free survival for patients receiving PDS01ADC + HAIP therapy was 12.7 months with a minimum follow-up of 13.1 months. Grade ≥3 toxicities occurred in 78% but were manageable and did not limit HAIP therapy. No patients developed a biliary stricture within 6 months of initiating treatment. Clinical responders to PDS01ADC demonstrated enhanced peripheral immune activation, including elevated levels of circulating T-cell subsets with stem-like features, and increased CD8 ⁺ T-cell:T regulatory cell ratio in tissue biopsies. CONCLUSION: Addition of PDS01ADC is not detrimental to HAIP therapy and is associated with both systemic and intratumoral immune modulation. Initial results warrant continuation to full enrollment for further evaluation of clinical and scientific end points.

Figure S11. ScRNA analysis of TILs derived memory CD4 T cells.

Figure S13. Flowcytometric cell surface marker analysis of KLM-1 pancreatic cancer cell line.

Figure S1. Differential expression levels of LILRs in healthy human PBMC derived Immune cells.

Figure S4. Flowcytometric gating strategy of healthy human PBMC derived monocytes and lymphocytes subsets.

Immune checkpoint blockade for

Table S2. List of contacts between HLA-B*44:05 and DX17.

Figure S9. Flowcytometric gating strategies of lymphocytes and monocytes from TILs of Colorectal Cancer patient.