Introduction

Invariant Natural Killer T (iNKT) cells are lipid-specific T lymphocytes endowed with antitumor activities and whose infiltration is considered a positive prognostic factor in colorectal cancer (CRC). In this context, we have previously demonstrated that, at steady state, iNKT cells eliminate human CRC cells by releasing perforin and granzyme B (1).
However, we reported that the tumor-associated pathobiont Fusobacterium nucleatum is capable of inducing IL17 and GM-CSF expression in iNKT cells, promoting iNKT-mediated recruitment of neutrophils with PMN-MDSCs- like phenotype and functions. Tumor co-infiltration by iNKT cells and neutrophils in a large cohort CRC patients of correlates with negative clinical outcomes (2). Since the gut microbiota is an oncogenic driver of CRC and intestinal microbes represent potent stimulators of iNKT cell responses we analyzed the tumor-associated mucosal microbiota of the 35 CRC patients by 16S rRNA gene sequencing, revealing the presence in a large fraction of patients of Porphyromonas gingivalis, an opportunistic oral pathobiont often associated with inflammatory diseases and cancers.

Aims & Methods

Very little is known about how the function of iNKT cells is influenced by Porphyromonas gingivalis. Our aim is to assess changes in human iNKT cell phenotype and function provoked by this bacterium, and how these changes might contribute to colon cancer progression.
Human iNKT cell phenotype, functional profile and cytotoxic capabilites were evaluated upon stimulation with P. gingivalis isolates in vitro. Subsequently, as validation, P. gingivalis was administered to AOM/DSS-treated mice and evaluated iNKT cell phenotype and tumor progression. P. gingivalis-exposed human iNKT cells were sequenced and candidate molecules were tested in vitro in killing assays with human CRC epithelial cells.

Results

We observed that, compared to cells stimulated with aGalCer (a prototypical iNKT cell antigen), iNKT cells primed with P. gingivalis showed lower levels of interferon gamma, together with impaired cytotoxicity against CRC cell lines. These cells were also characterized by an increased expression of GM-CSF, and interleukins -17 and -10, as well as an increased capability of neutrophil chemotaxis. This pro-tumorigenic phenotype was confirmed in AOM/DSS-treated mice, as P. gingivalis-treated animals showed tumors of greater size, together with an increase in IL-17+ and GM-CSF+ iNKT cells. Pg-primed iNKT cells showed an enriched neutrophil chemotaxis gene signature, which included the chemokines CXCL1, CXCL2, CXCL5, CXCL8, CCL2 and CCL4 and of proinflammatory proteins acting on the cytotoxic machinery of NKT cells inhibiting their killing functions.

Conclusion

These results demonstrate that P. gingivalis can promote colon tumorigenesis by acting on iNKT cells, both by hampering antitumor properties and promoting the release of pro-tumorigenic cytokines and chemokines, which further act on myelo-phagocytic cells.

References

1. Díaz-Basabe, A., et al. (2021). Human intestinal and circulating invariant natural killer T cells are cytotoxic against colorectal cancer cells via the perforin–granzyme pathway. Mol Oncol, 15: 3385-3403. https://doi.org/10.1002/1878-0261.13104

Introduction

With the increase in the opportunity to follow up patients with risk factors for pancreatic cancer, such as IPMN, using imaging tests, it is generally considered to discontinue follow-up for those who are elderly and not eligible for surgery. There are few studies comparing the group of patients who underwent surgery with those who did not undergo surgery among super-elderly individuals aged 80 years or older.

Aims & Methods

This was a retrospective, single-center study aimed at examining the extension of survival by surgery in healthy super-elderly patients aged 80 years and over with pancreatic cancer. Among the 183 patients diagnosed with histologically confirmed conventional ductal adenocarcinoma of the pancreas at our hospital between April 2011 and March 2021, we targeted 20 cases aged 80 years or older, whose clinical courses were available, and who were considered to be resectable or borderline resectable with an ECOG Performance Status of 0. Of the 20 cases, 11 underwent surgery and 9 did not, and we retrospectively compared the surgery group and non-surgery group.

Results

The age of the surgery group was 83±5 years, while that of the non-surgery group was 88±6 years, which was significantly higher in the non-surgery group. However, there was no significant difference in gender, symptom rate, abdominal pain rate, or blood test results for Alb, AMY, CEA, or CA19-9 levels. Tumor localization was pancreatic head (64%), pancreatic body (18%), and pancreatic tail (18%) in the surgery group, while all cases in the non-surgery group were located in the pancreatic head. All cases in the surgery group were Resectable on CT imaging, and they underwent pancreaticoduodenectomy (73%), distal pancreatectomy (18%), or total pancreatectomy (9%) without NeoAdjuvant Chemotherapy. In the non-surgery group, 89% were Resectable and 11% were Borderline on CT imaging, and the reasons for non-resection were age (45%), patient preference (33%), past medical history (11%), and surgeon's judgment (11%). Chemotherapy was administered in 64% of the surgery group and 67% of the non-surgery group, and second-line or later treatment was administered in 27% of the surgery group and 22% of the non-surgery group. In addition, radiation therapy was administered in 18% of the surgery group. The median survival time (IQR) was 333 (251-426.5) days in the surgery group and 226 (137-728) days in the non-surgery group, which was significantly longer in the surgery group. All cases in the surgery group died of the original disease, while 78% of the non-surgery group died of the original disease and 22% died of cerebral infarction.

Conclusion

Surgery is expected to prolong the survival period even in super-elderly individuals, and imaging follow-up for early detection of pancreatic cancer is important even in super-elderly individuals if they have a surgical tolerance.

Introduction

Microbiome analysis is a promising approach for the application of findings from basic research in the clinical field. However, there is an ongoing debate about the consistency of different bioinformatics analysis platforms. The lack of established standards affects the implementation of research findings.

Aims & Methods

In our study, we demonstrated that the results on the microbial composition of gastric mucosal biopsies are only slightly influenced by the efficacy of different microbiome analysis platforms. Furthermore, we investigated whether recent revisions to the International Code of Nomenclature for Prokaryotes and consequent alterations in taxonomic classification influenced the findings.
Gastric biopsy specimens were obtained from gastric cancer patients with well-defined clinical profiles (n=40), including individuals with and without Helicobacter pylori (H. pylori) infection, and from control subjects (n=39) with and without H. pylori infection. DNA extraction and 16S rRNA gene sequencing (V1-V2) was performed at one site, and the resulting fastQ files were distributed to five expert teams for independent analysis. Three widely used bioinformatics pipelines for microbiome analysis, namely DADA2, MOTHUR and QIIME2, were used for the analysis.

Results

The generated fastQ files exhibited no platform-dependent clustering, indicating the comparability of the sequences. Microbial diversity was slightly reduced in both GC groups and significantly reduced in the H. pylori positive controls. This is due to an extreme increase in the relative abundance of H. pylori in this group. It also follows that the samples of the H. pylori positive control group have a very high similarity and form the clearest cluster in the corresponding analysis. Regardless of the protocol used, H. pylori status, microbial diversity and relative bacterial abundance were reproducible on all platforms. Although minor differences in performance were detected and assessed, similarity analysis and bacterial enrichment assessment also yielded consistent results. The introduction of the new taxonomy only had a major impact on the Proteobacteria phylum, as it originally also included H. pylori according to the old taxonomy, but this is now assigned to the new Campilobacterota phylum.

Conclusion

In summary, our findings demonstrate that disparate microbiome analysis methodologies employed by independent research teams can yield comparable results from identical datasets.

Clinical Case Summary

In advanced chronic liver disease (ACLD) patients, the endoscopic evidence of esophageal and/or gastric varices denotes the presence of clinically significant portal hypertension (CPSH), representing the main clinical consequence of portal hypertension (PH).
In this setting, although infrequently, besides typical sites (i.e., esophagus and stomach), PH-related varices can manifest even in other atypical portosystemic-shunt districts, including the vagina. CSPH-related vaginal varices bleeding can occur as a rare complication documented by limited case reports.
A 62-year-old female patient with viral (eradicated Hepatitis C Virus infection)-related- decompensated (grade 2 ascites)-ACLD, complicated by CSPH (grade F2 esophageal varices), was admitted to our Department in February 2025 for microcitic anemia. Anamnesis revealed a previous gastric cancer managed with total gastrectomy resection in 2021.
On admission, abdominal imaging showed radiological signs of ACLD/PH (coarse pattern, splenomegaly, dilated splenic-portal axis). Esophagogastroduodenoscopy ruled out active sources of bleeding, whereas colonoscopy revealed anal congestive not-bleeding varices. During the hospitalization, an episode of metrorrhagia imposed gynecologic evaluation with vaginal speculum examination. This assessment documented vaginal varices with the relative rupture as the current cause of the bleeding, suggesting this site is the further district for (CS)PH manifestation consequently to potentially occurred total gastrectomy-determined changes in abdominal vascular anatomy.
In ACLD patients receiving surgery altering abdominal vascularization, variceal bleeding represents a possible CSPH-related complication occurring even in atypical sites, such as the vagina. This scenario imposes an adequate tailored management, monitoring unusual bleeding signs, and adopting appropriate preventive strategies by including gynecologic evaluation in follow-up.

References

1. McHugh PP, Jeon H, Gedaly R, Johnston TD, Depriest PD, Ranjan D. Vaginal varices with massive hemorrhage in a patient with nonalcoholic steatohepatitis and portal hypertension: Successful treatment with liver transplantation. Liver Transpl. 2008 Oct;14(10):1538-40. doi: 10.1002/lt.21506. PMID: 18825698.
2. Kreek MJ, Raziano JV, Hardy RE, Jeffries GH. Portal hypertension with bleeding vaginal varices. Ann Intern Med. 1967 Apr;66(4):756-9. doi: 10.7326/0003-4819-66-4-756. PMID: 6067181.
3. Chan G, Taqi A, Drage M, Quan D. A case report of massive vaginal variceal hemorrhage in a patient with cirrhosis secondary to nonalcoholic steatohepatitis unresolved even after successful liver transplantation. Transplant Proc. 2013 Mar;45(2):835-7. doi: 10.1016/j.transproceed.2012.04.033. Epub 2012 Sep 19. PMID: 23267803.
4. Lebrec D, Benhamou JP. Ectopic varices in portal hypertension. Clin Gastroenterol. 1985 Jan;14(1):105-21. PMID: 3872747.
5. Nagata J, et al. Vaginal bleeding in a patient with type C liver cirrhosis without a past history of laparotomy: successful treatment with partial splenic artery embolization. Clin J Gastroenterol. 2012 Aug;5(4):275-81. doi: 10.1007/s12328-012-0316-3. Epub 2012 Jun 30. PMID: 26182393.

Introduction

Detective flow imaging (DFI) is a novel technology that enables the evaluation of low-speed blood flow signal that is difficult to visualize with conventional Doppler techniques^1)-3). This technique allows for evaluating microvascular signals within tumors in endoscopic ultrasound (EUS) without using contrast agents. In this study, we aimed to investigate the usefulness of DFI for the diagnosis of biliary tract lesions.

Aims & Methods

A retrospective study was conducted of patients who underwent DFI with EUS between September 2019 and December 2022 at our institution. Patients with biliary tract lesions diagnosed by either endoscopy or pathology were included. Malignant biliary tract lesions were defined as those with histopathological diagnoses of non-invasive or invasive carcinoma. To evaluate the diagnostic ability of DFI for malignant biliary tract lesions, we assessed the presence of tortuous or variable caliber vessels in DFI findings. Inter-observer agreement and the diagnostic ability of DFI in identifying malignant biliary tract lesions were also assessed. Additionally, the DFI findings of intraductal papillary neoplasms (IPNB) and cholangiocarcinoma in patients who underwent surgical resection.

Results

There were 17 cases of cholangiocarcinoma, 9 cases of IPNB, 3 cases of bile duct stone, and 3 other cases. The final diagnosis of biliary tract lesion was made by surgery in 20 cases and by EUS-FNA or biopsy in 9 cases. In the first analysis, the sensitivity and specificity for the presence of tortuous or variable caliber vessels for the diagnosis of malignancy were 80% and 43% for tortuous vessels and 68% and 100% for variable caliber vessels, respectively. Interobserver agreement was k=0.63 for tortuous vessels and k=0.75 for variable caliber vessels. In the second analysis, DFI identified stalk vessels in 7 of 8 cases (87.5%) of IPNB and in only 1 of 10 cases (10%) of invasive cholangiocarcinoma. The rate of concomitant stalk vessels was significantly higher in IPNB than in invasive cholangiocarcinoma (P<0.01).

Conclusion

DFI is a simple and easy-to-use technique that allows for the evaluation of blood flow in biliary tract lesions without the use of contrast agents. It has the potential to be a useful tool for the differential diagnosis of malignant biliary tract lesions.

References

1. Yamashita Y, Yoshikawa T, Yamazaki H et al. A novel endoscopic ultrasonography imaging technique for depicting microcirculation in pancreatobiliary lesions without the need for contrast-enhancement: a prospective exploratory study. Diagnostics (Basel) 2021; 11: 2018.
2. Yamashita Y, Yoshikawa T, Kawaji Y et al. Novel endoscopic ultrasonography imaging technique for visualizing microcirculation without contrast enhancement in subepithelial lesions: prospective study. Dig Endosc 2021; 33: 955–961.
3. Miwa H, Sugimori K, Maeda S. Vessel images of gallbladder polypoid lesions on detective flow imaging endoscopic ultrasonography. Dig Endosc 2023. doi: 10.1111/den.14530 [Online ahead of print].

Introduction

Ulcerative colitis (UC) has increasingly been recognized as a progressive disease. However, the benefit of starting early biologic therapy (BT) has not been established, contrary to Crohn´s disease. We aimed to assess if early intervention improves UC outcomes, particularly in preventing disease progression.

Aims & Methods

We conducted a retrospective multicenter study (11 centers) including adult UC patients who initiated BT until 2019 (minimum follow-up [FU]: 5 years). Early intervention was defined as BT initiation within ≤12 months of diagnosis. The primary outcome was UC progression at 5 and 10 years, defined as a composite of proximal extension, colonic structural changes (stenosis/ tubularization) and dysplasia/cancer. Secondary outcomes included clinical endpoints: hospitalizations, colectomy and corticosteroid use. Multivariate analysis was performed, adjusted for disease severity, extent, and prior activity.

Results

A total of 236 patients were included, with 28.8% (n=68) initiating BT within ≤12 months (mean FU after BT initiation: 8.06 ± 4.12 years).
Early intervention was associated with a reduced risk of proximal extension after BT initiation (p=0.012). In multivariable Cox regression, later BT initiation was an independent predictor of colonic structural changes at 10 years (HR 8.54; 95% CI 1.15-62.63; p=0.036). Later BT initiation independently predicted the composite outcome of progression (23.2% vs 5.9%) at 5 years (HR 5.97; 95% CI 1.42-25.13; p=0.015) and 10 years (HR 4.58; 95% CI 1.41-14.88; p=0.011). No differences in clinical outcomes were observed.
Longer disease activity before BT was also a predictor of the primary outcome at 5 years (HR 4.20; 95% CI 1.27–13.82; p=0.018) and 10 years (HR 3.60; 95% CI 1.28–10.12; p=0.015).

Conclusion

Starting BT early in the disease course may reduce the long-term risk of disease progression, reshaping the window of opportunity in UC.

References

Lujan, Rona, et al. "Early initiation of biologics and disease outcomes in adults and children with inflammatory bowel diseases: results from the Epidemiology Group of the Nationwide Israeli Inflammatory Bowel Disease Research Nucleus Cohort." Gastroenterology 166.5 (2024): 815-825.