Abstract Background Colorectal liver metastases (CRLM) are a major cause of cancer-related deaths. Glucose transporter 1 (GLUT1) is a key mediator of glycolytic metabolism in malignant and immune cells; however, the prognostic relevance of compartment-specific GLUT1 patterns in CRLM remains undefined. We hypothesized that spatial GLUT1 expression at the tumor–liver interface may reflect clinically relevant microenvironmental biology. Methods We retrospectively analyzed data of 192 patients who underwent curative-intent resection for CRLM (75 solitary; 117 multiple). GLUT1 expression was assessed by immunohistochemistry in the tumor tissue (Tu) and infiltration margin (Im) and was correlated with overall survival using Cox regression. Double immunofluorescence for CD8 and GLUT1 was performed for qualitative visualization at the infiltration margin. In an exploratory subset (n = 5), flow cytometry and in vitro killing assays were conducted on GLUT1⁺ versus GLUT1⁻ CD8⁺ tumor-infiltrating lymphocytes (TILs). Results Tumoral GLUT1 correlated with Ki67 (Spearman’s ρ = 0.31, p = 0.003) but was not independently associated with overall survival in the main cohort (multivariable HR 1.183, 95% CI 0.74–1.90; p = 0.485). In the main cohort, capsule presence remained strongly associated with improved survival (HR 0.35, 95% CI 0.21–0.58; p < 0.001). In contrast, high invasive-margin GLUT1 was independently associated with improved survival in the predefined solitary cohort (HR 0.379, 95% CI 0.18–0.81; p = 0.012). Double immunofluorescence demonstrated the qualitative co-localization of the GLUT1 signal with CD8⁺ cells at the infiltration margin. In exploratory assays (n = 5), flow cytometry suggested GLUT1 enrichment in CD8⁺ terminally differentiated effector memory T cells re-expressing CD45RA (TEMRA), and GLUT1⁺ TIL fractions showed higher in vitro tumor cell killing compared with GLUT1⁻ cells. Conclusion GLUT1 shows compartment- and context-dependent associations in the CRLM. While tumor-core GLUT1 is linked to proliferative activity, invasive-margin GLUT1 is associated with favorable outcomes in solitary metastases. Immunofluorescence and functional data provide hypothesis-generating context and warrant validation with quantitative spatial immune profiling and independent cohorts.

Der intraoperative Ultraschall (IOUS), ggf. als „contrast-enhanced intraoperative ultrasound“ (CE-IOUS), ist ein vielseitiges Instrument der chirurgischen Entscheidungsfindung. Er verbessert die intraoperative Orientierung, ermöglicht eine präzise Resektionsführung und trägt zur Vermeidung von Komplikationen und Folgeeingriffen bei. In Deutschland wird IOUS bislang jedoch nur selektiv eingesetzt – häufig durch nicht-operativ tätige Disziplinen. Dies ist in bestimmten Konstellationen sinnvoll, stößt jedoch an Grenzen, sobald die Bildgebung unmittelbar in die chirurgische Handlung einfließt. Der klinische Nutzen des IOUS ist in mehreren Fachgebieten – insbesondere der Leberchirurgie, Neurochirurgie und operativen Senologie – gut belegt. Dennoch fehlt bislang eine systematische Verankerung in der chirurgischen Ausbildung, im klinischen Alltag und in den Leitlinien. In der Viszeralchirurgie verfügen bislang nur wenige Operateure über eigene IOUS-Erfahrung. In anderen Bereichen wie der Gefäß- oder Transplantationschirurgie gehört der Einsatz bereits zum Standard, jedoch ohne dass dieser durch randomisierte Studien abgesichert wäre. Die DEGUM als führende Fachgesellschaft für Ultraschall in der Medizin spricht sich daher für eine breitere und strukturierte Integration des IOUS in der operativen Medizin aus. Sie bietet praxisnahe Weiterbildungsformate in Modulform an, fördert wissenschaftliche Projekte und setzt sich in der Leitlinienarbeit dafür ein, bestehende Evidenz sichtbar und zukünftige nutzbar zu machen. Ziel ist es, IOUS als festen Bestandteil qualitätsgesicherter chirurgischer Versorgung zu etablieren – interdisziplinär kooperativ, aber mit eigener sonographischer Kompetenz auf operativer Seite. Daher empfiehlt die DEGUM die Anwendung von IOUS zur Kontrolle und Steuerung bei Tumorresektionen, zur Navigation bei anatomisch unklaren Situs, zur Beurteilung von Gefäßanastomosen bzw. der Organperfusion und zur erweiterten Diagnostik, insbesondere in der Leberchirurgie, ggf. unter ergänzendem Einsatz von Kontrastmittelsonographie.

Patient-derived xenografts (PDXs) provide biologically relevant models and potential platforms for the development of treatment strategies for precision medicine in pancreatic cancer. Furthermore, circulating epithelial tumor cells (CETCs/CTCs) are released into the bloodstream by solid tumors and a rare subpopulation—circulating cancer stem cells (cCSCs) – is considered to be responsible for recurrence and plays a key role in metastasis. For the identification of cCSCs, an innovative in vitro assay to generate tumorspheres was established in this study. The number of tumorspheres and CETCs/CTCs was analyzed perioperatively in 25 pancreatic cancer patients. Additionally, an individual in vivo chorioallantoic membrane (CAM) culture system was used to generate PDXs from these tumorspheres. While overall correlations of CETCs/CTCs with clinicopathological parameters did not reach statistical significance, a significant difference in the number of tumorspheres was observed between patient subgroups with lower and higher UICC stages. This finding underscores their potential as biomarkers, providing valuable insights into clinical decision-making and tumor progression. The application of tumorspheres on the CAM successfully established PDXs within 7 days. These xenografts closely resembled the histological features of the primary tumor. Hence, this model represents a novel and fast option for individualized testing of new therapies for PDAC.

Background: Portal vein thrombosis (PVT) leads to portal hypertension (PH) with its sequelae. Computed tomography spleno-mesenterico-portography (CT-SMPG) combines sequential CT spleno-portography and CT mesenterico-portography. CT-SMPG comprehensively illustrates the venous hemodynamic changes due to PH. Objective: To assess the effects of PV confluence thrombosis (PVCT) and liver cirrhosis on venous blood flow characteristics of patients with PVT. Method: CT-SMPG was performed in 21 patients with chronic PVT. CT-SMPG was compared to standard contrast-enhanced CT (CECT) and gastroscopy concerning the patency of splanchnic veins, varices and venous congestion. Results: PVCT had a significant effect on perfusion patterns: in patients without PVCT, esophageal varices (EV) and gastric varices were supplied by either the splenic vein (SV), the superior mesenteric vein (SMV), or both. In patients with PVCT, EV and gastric varices were mostly supplied by the SV (p = 0.021, p = 0.016). In patients without PVCT, small bowel varices were fed by both systems or the SMV, while in patients with PVCT they were fed by the SMV (p = 0.031). No statistically significant changes were detected regarding gastropathy, colorectal varices and small bowel congestion. Liver cirrhosis had no statistically relevant effect on hemodynamics. Conclusions: In CT-SMPG, patients with PVCT showed different venous hemodynamics to patients without PVCT, and this can serve as a basis for selecting therapy options.

Intraoperative ultrasound (IOUS), also potentially as contrast-enhanced (CE)-IOUS, is a versatile tool for surgical decision-making. It enhances intraoperative orientation, enables precise resection guidance and helps prevent complications and follow-up procedures; however, in Germany, IOUS is currently only selectively used, often in nonsurgical disciplines. While this is reasonable in certain scenarios, it reaches its limits when imaging directly influences surgical actions. The clinical benefits of IOUS are well-documented in several specialties, particularly liver surgery, neurosurgery and surgical senology. Nevertheless, a systematic integration into surgical training, clinical practice, and guidelines is still lacking. In visceral surgery only a few surgeons have their own IOUS experience. In other fields, such as vascular or transplantation surgery, its use is already standard, although this has not been confirmed by randomized studies. The German Society for Ultrasound in Medicine (DEGUM), as the leading professional society for ultrasound in medicine, advocates a broader and structured integration of IOUS into surgical medicine. It provides practical modular training formats, promotes scientific projects and works with guidelines to make existing evidence visible and future evidence usable. The goal is to establish IOUS as a fixed component of quality-assured surgical care both interdisciplinary and cooperative but with independent sonographic expertise on the surgical side. Therefore, DEGUM recommends the use of IOUS for control and guidance during tumor resections, for navigation in anatomically unclear situations, for assessment of vascular anastomoses or organ perfusion and for extended diagnostics, particularly in liver surgery, potentially with the supplementary use of contrast-enhanced ultrasound.

The prognosis for patients diagnosed with cholangiocarcinoma (CCA) is dismal, with an overall 5-year-mortality rate of 80%. Therapeutic approaches for this cancer are very limited and the only curative treatment is total surgical resection despite recent advancements in CCA research. However, only a minority of patients are eligible for surgery due to late-stage diagnosis. Therefore, there is an urgent need to gain a deeper understanding of CCA and to discover new treatments, which can be achieved by utilization and optimization of 3D tumor models. Traditional 2D cell culture is still undeniably important in cancer research, especially for the discovery of biomarkers and drug screening. However, classical 2D tumor models do not represent the tumor biology in its full complexity as they lack the vital interactions between cancer cells, angiogenesis, and tumor microenvironment. In recent years, 3D models, including spheroids, 3D co-culture systems, organoids, tumors-on-a-chip, and the in vivo chorioallantoic membrane (CAM) model, have been used for CCA research. These models enable the study of the tumor microenvironment, investigation of metastases, drug development and testing, cholangiocarcinogenesis and personalized therapy. This review summarizes the applications of the different 3D tumor models that have been used for the investigation of CCA. Moreover, the advantages and disadvantages of the different 3D tumor models are discussed, and suggestions for future research possibilities are described. By optimizing 3D models, the gap between basic research findings and clinical applications can be bridged, enabling the discovery of more effective therapies for CCA and other cancers.

Abstract Background: Pancreatic cancer carries a poor prognosis in the majority of patients, with an overall 5-year survival rate of less than 10%. Therefore, the development of new therapies and preclinical models is crucial and urgently needed. Circulating cancer stem cells (cCSCs) are a subset of circulating cancer cells that have stem cell-like properties and play a significant role in tumor progression, metastasis, and treatment resistance. Patient-derived xenografts (PDX) reflect the genotypic and phenotypic landscape of patients' cancers and are useful for drug screening, biomarker development, and preclinical evaluation of personalized medicine strategies. In this study, we describe a fast, simple, and cost effective method to generate PDX on Chorioallantoic Membrane (CAM) from circulating cancer stem cells. Methods: In this study, primary cultures from circulating cancer stem cells were established using sphere-forming assays from 30 patients with newly diagnosed pancreatic cancer. Subsequently, tumorspheres were transplanted onto the CAM membrane of fertilized chicken eggs to form patient- derived xenografts. Results: For the identification of cCSCs from peripheral blood of pancreatic cancer patients, an innovative in vitro sphere-forming assay was established. The number of tumorspheres was correlated with clinicopathological parameters like tumor size, lymph node involvement and grading status of the primary tumor. Implantation of tumorspheres onto the CAM was successful in ~90 % of the applied samples. The histological analysis suggests that the PDX tumors were morphologically and pathologically identical to primary tumor tissue. Conclusions: The number of tumorspheres was associated with clinicopathological parameters. Further follow-up is needed to evaluate the prognostic significance of tumorsphere detection in pancreatic cancer. Generation of PDX from cCSCs on CAM has a high success rate and may become an integral part of the drug development arena, including drug screening and biomarker development. Citation Format: Monika Pizon, Dorothea Schott, Andreas Ettner-Sittner, Benedikt J. Wagner, Katharina Pachmann, Thiha Aung, Christina Hackl, Silke Haerteis. Patient-derived xenografts from circulating cancer stem cells and their potential utility in personalized medicine of pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6900.

Ultra high frequency (UHF) ultrasound enables the visualization of very small structures that cannot be detected by conventional ultrasound. The utilization of UHF imaging as a new imaging technique for the 3D-in-vivo chorioallantoic membrane (CAM) model can facilitate new insights into tissue perfusion and survival. Therefore, human renal cystic tissue was grafted onto the CAM and examined using UHF ultrasound imaging. Due to the unprecedented resolution of UHF ultrasound, it was possible to visualize microvessels, their development, and the formation of anastomoses. This enabled the observation of anastomoses between human and chicken vessels only 12 h after transplantation. These observations were validated by 3D reconstructions from a light sheet microscopy image stack, indocyanine green angiography, and histological analysis. Contrary to the assumption that the nutrient supply of the human cystic tissue and the gas exchange happens through diffusion from CAM vessels, this study shows that the vasculature of the human cystic tissue is directly connected to the blood vessels of the CAM and perfusion is established within a short period. Therefore, this in-vivo model combined with UHF imaging appears to be the ideal platform for studying the effects of intravenously applied therapeutics to inhibit renal cyst growth.

BACKGROUND/AIM:The aim of this study was to describe and evaluate the patterns, perioperative outcomes, and survival rates of patients subjected to hepatic resections for ovarian-derived liver metastasis as part of cytoreductive surgery with or without hyperthermic intraperitoneal chemotherapy (HIPEC). Furthermore, we investigated two subgroups of tumor patterns: hematogenous liver metastasis and infiltrative liver metastatic spread.PATIENTS AND METHODS:A retrospective study was conducted. Patients from a University Tertiary Hepatic and Peritoneal Surface Malignancy Center with primary or recurrent ovarian cancer, who underwent liver resection as part of cytoreductive surgery between January 1992 and December 2022, were included.RESULTS:Data from 35 patients were analyzed. Both median overall survival (OS) and disease-specific survival (DSS) were 24.97 months. In a multivariate setting, the combined effect of age, peritoneal carcinomatosis index, body mass index, hematogenous liver metastasis vs. infiltrative spread types, and HIPEC (HR=0.2372; 95%CI=0.0719-0.7823; p=0.0181) over OS was tested. Survival analysis revealed no differences between the two metastatic spread types (OS: p=0.9720; DSS: p=0.9610). Younger age (p=0.0301), splenectomy (p=0.0320), lesser omentectomy (p=0.0178), and right upper quadrant peritonectomy (p=0.0373) were more characteristic for those patients with infiltrative liver metastatic spread.CONCLUSION:Complete cytoreductive surgery, including hepatic resection is a feasible approach with or without additional HIPEC, which may provide survival benefit for patients with advanced and/or recurrent ovarian cancer. If metastatic and infiltrative liver involvement is suspected, liver-specific imaging is recommended.

Although significant improvements have been made in the treatment of pancreatic cancer, its prognosis remains poor with an overall 5-year survival rate of less than 10%. New experimental approaches are necessary to develop novel therapeutics. In this study, the investigation of pancreatic cancer tissue growth in the chorioallantoic membrane (CAM) model and the subsequent use of indocyanine green (ICG) injections for the verification of intratumoral perfusion was conducted. ICG was injected into the CAM vasculature to visualize the perfusion of the tumor tissue. The presence of metastasis was investigated through PCR for the human-specific ALU element in the liver of the chicken embryo. Additionally, the usage of cryopreserved pancreatic tumors was established. Intratumoral perfusion of tumor tissue on the CAM was observed in recently obtained and cryopreserved tumors. ALU-PCR detected metastasis in the chick embryos’ livers. After cryopreservation, the tissue was still vital, and the xenografts generated from these tumors resembled the histological features of the primary tumor. This methodology represents the proof of principle for intravenous drug testing of pancreatic cancer in the CAM model. The cryopreserved tumors can be used for testing novel therapeutics and can be integrated into the molecular tumor board, facilitating personalized tumor treatment.

PURPOSE:Complete resection of the affected tissue remains the best curative treatment option for liver-derived tumors and colorectal liver metastases. In addition to preoperative cross-sectional imaging, contrast-enhanced intraoperative ultrasound (CE-IOUS) plays a crucial role in the detection and localization of all liver lesions. However, its exact role is unclear. This study was designed to evaluate the clinical and oncological impact of using CE-IOUS in the surgical treatment of these diseases.MATERIALS AND METHODS:Over the three-year study period, 206 patients with primary liver tumors and hepatic metastases were enrolled in this prospective, monocentric study to evaluate the impact of CE-IOUS in liver surgery. Secondary outcomes included comparing the sensitivity and specificity of CE-IOUS with existing preoperative imaging modalities and identifying preoperative parameters that could predict a strategic impact of CE-IOUS. In addition, the oncological significance of CE-IOUS was evaluated using a case-cohort design with a minimum follow-up of 18 months.RESULTS:CE-IOUS findings led to a change in surgical strategy in 34% of cases (n=70/206). The accuracy in cases with a major change could be confirmed histopathologically in 71.4% of cases (n=25/35). The impact could not be predicted using parameters assumed to be clinically relevant. An oncological benefit of a CE-IOUS adapted surgical approach was demonstrated in patients suffering from HCC and colorectal liver metastases.CONCLUSION:CE-IOUS may significantly increase R0 resection rates and should therefore be used routinely as an additional staging method, especially in complex liver surgery.

Supplementary Data from Up-Regulation of Krüppel-Like Factor 5 in Pancreatic Cancer Is Promoted by Interleukin-1β Signaling and Hypoxia-Inducible Factor-1α

Supplementary Fig. S3 from Dual targeting of Raf and VEGF receptor 2 reduces growth and metastasis of pancreatic cancer through direct effects on tumor cells, endothelial cells, and pericytes

Although there have been improvements in the treatment of pancreatic cancer in recent years, the prognosis for this disease remains poor, with an overall 5-year survival rate of less than 10%. Therefore, new therapies and preclinical models are urgently needed. Circulating cancer cells, and in particular their very rare subpopulation, circulating cancer stem cells (cCSC) are good candidates for generating preclinical models, making it possible to follow up the spatial and temporal heterogeneity of tumor tissues. This method is a non-invasive liquid biopsy that can be obtained at any stage of the disease. In this study, we sought to set up a novel model in which patient-derived xenograft can be generated on Chorioallantoic Membrane (CAM) from circulating cancer stem cells. In this study, primary cultures from circulating cancer stem cells were established using sphere-forming assays from 30 patients with newly diagnosed pancreatic cancer. Subsequently, tumorspheres were transplanted onto the CAM membrane of fertilized chicken eggs to form secondary microtumors. Herein, we report an innovative in vitro platform for cultivation of cCSCs from peripheral blood of pancreatic cancer patients. The number of tumorspheres increased significantly with tumor size, lymph node involvement and grading status of the primary tumor. Implantation of tumorspheres onto the CAM was successful in ∼90 % of the applied samples. The first histological analysis suggests that the main histopathological features closely resemble the original pancreatic tumors. The number of tumorspheres is associated with some clinicopathological parameters, but further follow-up is needed to evaluate the prognostic significance of tumorsphere detection in pancreatic cancer. PDXs generated from cCSCs on CAM membrane offer a promising approach for personalized medicine in pancreatic cancer, providing a platform for the study of individual patient tumors and the development of more effective treatments.