INTRODUCTION:The identification of risk factors for precursor lesions of colorectal cancer (CRC) holds great promise in the context of prevention. With this study, we aimed to identify patient characteristics associated with colorectal polyps (CPs) and polyp features of potential malignant progression. Furthermore, a potential association with gut microbiota in this context was investigated. METHODS:In this single-center study, a total of 162 patients with CPs and 91 control patients were included. Multiple variables including information on lifestyle, diet, serum parameters, and gut microbiota, analyzed by 16S-rRNA gene amplicon sequencing and functional imputations (Picrust2), were related to different aspects of CPs. RESULTS:We observed that elevated serum alkaline phosphatase (AP) levels were significantly associated with the presence of high-grade dysplastic polyps. This association was further seen for patients with CRC. Thereby, AP correlated with other parameters of liver function. We did not observe significant changes in the gut microbiota between patients with CP and their respective controls. However, a trend toward a lower alpha-diversity was seen in patients with CRC. Interestingly, AP was identified as a possible clinical effect modifier of stool sample beta diversity. DISCUSSION:We show for the first time an increased AP in premalignant CP. Furthermore, AP showed a significant influence on the microbial composition of the intestine. Relatively elevated liver enzymes, especially AP, may contribute to the detection of precancerous dysplastic or neoplastic changes in colorectal lesions. The association between elevated AP, premalignant CP, and the microbiome merits further study.

Zusammenfassung Präventionsstrategien bei Ösophaguskarzinomen Den Schlüsselfaktor gastrointestinaler Tumorprävention bildet die Behebung inflammatorischer Einflüsse, welche sich für die 2 ösophagealen Tumorentitäten Plattenepithel- und Adenokarzinom unterscheiden. Der steigenden Inzidenz der Adenokarzinome sollte durch Therapie von Übergewicht und gastroösophagealer Refluxkrankheit begegnet werden. Ein endoskopisches Screening ist in Deutschland nicht etabliert, hingegen erfolgt bei Barrett-Metaplasie eine engmaschige Surveillance. Für den gezielten Einsatz der Endoskopie ist zukünftig eine bessere Stratifizierung von Risikopatienten vonnöten; entsprechende Prädiktionsmodelle sind Gegenstand aktueller Forschung. Therapie der Plattenepithelkarzinome des Ösophagus Neben endoskopischen Resektionsoptionen sehr früher Befunde besteht die Hauptneuerung der letzten Jahre im prognoseverbessernden Einsatz von Immun-Checkpoint-Inhibitoren in der adjuvanten und palliativen Systemtherapie. Der PD-L1-Status dient hierbei als stratifizierender Biomarker. Therapie der Adenokarzinome des Ösophagus und des ösophagogastralen Übergangs Aufgrund ihrer engen molekularbiologischen Verwandtschaft erfolgt die Systemtherapie der AEG-Tumore analog der Magenkarzinome. In der perioperativen Therapie lokal fortgeschrittener Befunde hat jüngst die Chemotherapie nach dem FLOT-Protokoll ihren Stellenwert weiter unterstreichen können. In metastasierten Stadien wird die Chemotherapie biomarkerstratifiziert durch Checkpoint-Inhibitoren sowie Antikörper gegen Her2 und fortan auch Claudin-18.2 ergänzt. Mit dem Antikörper-Wirkstoff-Konjugat Trastuzumab-Deruxtecan wurde eine Her2-gerichtete Zweitlinienoption etabliert.

Supplementary Table S2. Tissue Microarray Metadata (merged data from 4 TMAs used in the study): Impact of stage, T-stage, N-stage, and tumor grade on c-MET positivity and H-score of EAC samples.

Supplementary Figure S5. Gross anatomy of mouse stomach.

Ösophageale Tumoren haben unter den gastrointestinalen Malignomen eine schlechte Prognose. Obwohl weltweit das Plattenepithelkarzinom des Ösophagus (ESCC) dominiert, ist in den westlichen Ländern eine deutliche Zunahme des Adenokarzinoms (EAC) zu beobachten. Das EAC ist meist am Übergang vom Ösophagus zum Magen lokalisiert und kann sich aus dem Barrett-Ösophagus (BE), einer Folge der chronischen Refluxkrankheit, entwickeln. Das ESCC hingegen tritt hauptsächlich im oberen Drittel der Speiseröhre auf und wird durch Zigaretten- und Alkoholkonsum begünstigt. Die Unterschiede zwischen EAC und ESCC gehen weit über die Risikofaktoren hinaus, aber werden trotzdem unter dem Oberbegriff „Ösophaguskarzinom“ zusammengefasst. Um den aktuellen Stand der Wissenschaft , v. a. hinsichtlich der Unterschiede zwischen EAC und ESCC sowie der Karzinogenese des EAC, zu ermitteln, wurde eine selektive Literaturrecherche in der Datenbank PubMed durchgeführt. Die Suchbegriffe umfassten u. a.: Ösophaguskarzinom, Risikofaktoren für EAC und ESCC, BE, Tumormikromilieu (TME) und die Rolle des Immunsystems. Es wurde Artikel bevorzugt, die zwischen EAC und ESCC unterscheiden. In den vergangenen Jahren haben mehrere Studien darauf hingedeutet, dass das EAC von einer Vorläuferzelle des Magens abstammt. Es wurde zudem festgestellt, dass das EAC die Eigenschaften von Magentumoren aufweist, während das ESCC eher dem Kopf-Hals-Tumor ähnelt. Die Karzinogenese des EAC ist ein komplexer Prozess, der durch die Interaktion verschiedener Faktoren auf zellulärer, immunologischer und tumormikrobieller Ebene gekennzeichnet ist. Einige Funktionen von Zelltypen wie tumorassoziierten Makrophagen, regulatorischen T‑Zellen und T‑Helferzellen wurden bereits identifiziert. Dennoch besteht weiterhin Forschungsbedarf, um die Karzinogenese des EAC vollständig zu verstehen.

Ex vivo fluorescence imaging of L2-IL1β mice: A, Representative white light and fluorescence images of the excised stomach taken from L2-IL1β mice. The white dotted line shows the SCJ, which is within the stomach in mice. A protruding, irregular lesion is visible at the EGJ and SCJ in the stomach taken from a high-score group mouse, with the corresponding fluorescence image below. B, Upregulation of c-MET in dysplastic lesions from the low-, intermediate-, and high-score groups. C, Correlation between dysplasia score and level of c-MET expression. D, Box-whisker plot of ROI analysis, which shows increasing fluorescence from low- to intermediate- to high-score lesions. E, Quantification of TBR in ex vivo IVIS fluorescence imaging. Range of box-whisker plots indicate minimum to maximum. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. Eso, esophagus; H&E, hematoxylin and eosin; Max, maximum; Min, minimum; ns, not significant; Sbkg, signal intensities of the background; SJC, squamocolumnar junction; Slesion, signal intensities of the lesion.

Supplementary Figure S4. Fluorescence molecular endoscopy study schema.

BACKGROUND:While p53 mutations occur early in Barrett's oesophagus (BE) progression to oesophageal adenocarcinoma (EAC), their role in gastric cardia stem cells remains unclear. OBJECTIVE:This study investigates the impact of p53 mutation on the fate and function of cardia progenitor cells in BE to EAC progression, particularly under the duress of chronic injury. DESIGN:We used a BE mouse model (L2-IL1β) harbouring a Trp53 mutation (R172H) to study the effects of p53 on Cck2r+ cardia progenitor cells. We employed lineage tracing, pathological analysis, organoid cultures, single-cell RNA sequencing (scRNA-seq) and computational analyses to investigate changes in progenitor cell behaviour, differentiation patterns and tumour progression. Additionally, we performed orthotopic transplantation of sorted metaplastic and mutant progenitor cells to assess their tumourigenic potential in vivo. RESULTS:The p53 mutation acts as a switch to expand progenitor cells and inhibit their differentiation towards metaplasia, but only amidst chronic injury. In L2-IL1β mice, p53 mutation increased progenitors expansion and lineage-tracing with a shift from metaplasia to dysplasia. scRNA-seq revealed dysplastic cells arise directly from mutant progenitors rather than progressing through metaplasia. In vitro, p53 mutation enhanced BE progenitors' organoid-forming efficiency, growth, DNA damage resistance and progression to aneuploidy. Sorted metaplastic cells grew poorly with no progression to dysplasia, while mutant progenitors gave rise to dysplasia in orthotopic transplantation. Computational analyses indicated that p53 mutation inhibited stem cell differentiation through Notch activation. CONCLUSIONS:p53 mutation contributes to BE progression by increasing expansion and fitness of undifferentiated cardia progenitors and preventing their differentiation towards metaplasia.

Supplementary Figure S3. Representative time course fluorescence imaging of the dual xenograft mouse model after EMI-137 injection.

Bile acids are a major component of gastro-esophageal refluxate, thought to contribute to the development of Barrett’s esophagus (BE) and esophageal adenocarcinoma (EAC). As the microbiome shifts with EAC progression and bile acids influence bacterial composition, we examined these connections in a multi-center, cross-sectional study. We analyzed biospecimens from patients undergoing endoscopy using LC-MS to quantify bile acids in gastric aspirates, 16S rRNA sequencing for tissue microbiome profiling, and RNA sequencing on BE or cardia tissue. Among 153 patients (52 controls, 101 BE: 50 no dysplasia, 10 indefinite, 17 low-grade dysplasia, 17 high-grade dysplasia, and 7 EAC), we observed increased Streptococcus in BE tissue; dysplasia and EAC were associated with more Lactobacillus and decreased Actinomyces and other genera. Refluxate bile acids were mainly conjugated, indicating minimal bacterial metabolism, while BE patients had elevated secondary bile acid levels. Streptococcus correlated with upregulation of IL6, FGF2, and HGF, and decreased Actinomyces showed the most associations with gene expression, including the oxidative phosphorylation pathway. We identified two distinct BE gene expression clusters independent of histology, bile acid, or microbiome composition. These findings suggest bile acids shape the BE microbiome and associate with gene expression changes potentially relevant to EAC development.

Supplementary Information S1. Details of the Fluorescence Molecular Endoscopy (FME) System; Representative videos with screenshot of FME in L2-IL1b transgenic mice

Innerhalb der letzten Jahrzehnte konnten in der (frühzeitigen) Diagnostik und Therapie sowie in der Primär-, Sekundär- und Tertiärprävention gastrointestinaler onkologischer Erkrankungen maßgebliche Fortschritte verzeichnet werden. Im Folgenden werden aktuelle Erkenntnisse und Empfehlungen in der Prävention von gastrointestinalen Tumoren zur Integration in den klinischen Praxisalltag zusammengefasst.

Locally advanced esophageal adenocarcinoma remains difficult to treat and the ecological and evolutionary dynamics responsible for resistance and recurrence are incompletely understood. Here, we performed longitudinal multiomic analysis of patients with esophageal adenocarcinoma in the MEMORI trial. Multi-region multi-timepoint whole-exome and paired transcriptome sequencing was performed on 27 patients before, during and after neoadjuvant treatment. We found major transcriptomic changes during treatment with upregulation of immune, stromal and oncogenic pathways. Genetic data revealed that clonal sweeps through treatment were rare. Imaging mass cytometry and T cell receptor sequencing revealed remodeling of the tumor microenvironment during treatment. The presence of genetic immune escape, a less-cytotoxic T cell phenotype and a lack of clonal T cell expansions were linked to poor treatment response. In summary, there were widespread transcriptional and environmental changes through treatment, with limited clonal replacement, suggestive of phenotypic plasticity.

AbstractPurpose: Esophageal cancer carries a poor prognosis with a 5-year overall survival of less than 20%. Barrett’s esophagus increases the risk of esophageal adenocarcinoma. The aim of this study was to investigate the ability of EMI-137, a mesenchymal–epithelial transition factor (c-MET)-targeting optical imaging tracer, to detect dysplasia in Barrett’s esophagus. Experimental Design: c-MET expression in human esophageal tissue was investigated using Gene Expression Omnibus datasets, tissue microarrays, and Barrett’s esophagus biopsies. EMI-137 was tested in a dual xenograft mouse model bearing OE33 (c-MET high expression) and FLO-1 (c-MET low expression) tumors. Fluorescence molecular endoscopy was performed in a mouse model of Barrett’s-like metaplasia and dysplasia (L2-IL1β). Tumors and organs of interest were evaluated through ex vivo fluorescence imaging. Results: MET mRNA expression analyses and c-MET immunostaining confirmed upregulation of c-MET in Barrett’s esophagus and esophageal adenocarcinoma compared with normal epithelium. There was strong accumulation of EMI-137 in OE33 xenografts 3 hours after injection, decreasing by more than 50% on coinjection of a 10-fold molar excess of unlabeled EMI-137. The target-to-background ratio at 3 hours after injection for OE33 and FLO-1 tumors was 10.08 and 1.42, respectively. Fluorescence molecular endoscopy of L2-IL1β mice showed uptake of EMI-137 in dysplastic lesions within Barrett’s esophagus with a target-to-background ratio of 1.9 in vivo and greater than 2 in ex vivo fluorescence imaging. Conclusions: EMI-137 accumulates in dysplastic lesions within Barrett’s esophagus and also in c-MET–positive esophageal adenocarcinoma. EMI-137 imaging has potential as a screening and surveillance tool for patients with Barrett’s esophagus and as a means to detecting dysplasia and esophageal adenocarcinoma.

Details of L2-IL1β ex vivo IVIS experiment. Average radiant efficiencies of ROIs were calculated from ex vivo IVIS images: left and right squamocolumnar junctions, the junction between the forestomach and the cardiac/corpus of the stomach, and EGJ, with the corresponding background regions located in the corpus of the stomach.