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IMMUNOREACT 19: IN RECTAL CANCER PATIENTS, PREVIOUS APPENDECTOMY IS ASSOCIATED TO AN ALTERED ANTIGEN PRESENTATION IN THE NORMAL MUCOSA

Anna Matteazzi 1, Melania Scarpa 2, Andromachi Kotsafti 3, Astghik Stepanyan 4, Gaia Tussardi 1, Antonio Rosato 2, Ottavia De Simoni 2, Francesca Bergamo 2, Valerio Pellegrini 5, Ivana Cataldo 5, Ceccon Carlotta 5, Anna Pozza 5, Roberta Salmaso 6, Federico Scognamiglio 4, Cesare Ruffolo 1, Silvia Negro 4, Isacco Maretto 1, Imerio Angriman 6, Chiara Vignotto 1, Luca Facci 1, Giorgio Rivella 1, Quoc Riccardo Bao 1, Andrea Baldo 1, Giulia Pozza 1, Marco Agostini 1, Maurizio Zizzo 7, Beatrice Salmaso 8, Dario Parini 8, GiovannI Pirozzolo 9, Giovanni Bordignon 10, Roberto Merenda 10, Laurino Licia 10, Isabella Mondi 10, Silvio Guerriero 11, Giuseppe Portale 12, Lavinia Ceccarini 12, Michele Antoniutti 12, Laura Marinelli 13, Mattia Barbareschi 13, Giovanni Bertalot 13, Alberto Brolese 13, Giulia Noaro 9, Giulia Becherucci 14, Andrea Porzionato 6, Francesco Cavallin 15, Barbara Di Camillo 6, Gaya Spolverato 1, Ignazio CASTAGLIUOLO 1, Matteo Fassan 5, Marco Scarpa 1

Affiliations

1 Azienda Ospedaliera Universitaria di Padova, Padova, Italy

2 Veneto Institute of Oncology IOV-IRCCS, Padova, Italy

3 Veneto Institute of Oncology (IOV-IRCCS), Padova, Italy

4 Azienda Ospedaliera Universitaria di Padova, Yerevan, Italy

5 Azienda ULSS n 2 Marca Trevigiana, Treviso, Italy

6 University of Padova, Padova, Italy

7 Arcispedale Santa Maria Nuova IRCCS, Reggio Emilia, Italy

8 Azienda ULSS n 5 Polesana, Rovigo, Italy

9 Azienda ULSS n 6 Euganea, Padova, Italy

10 Azienda ULSS n 3 Serenissima, Venezia, Italy

11 Ospedale di Fermo, ASUR 4, Fermo, Italy

12 Azienda ULSS n 7 Pedemontana, Bassano del Grappa, Italy

13 Azienda Provinciale per i Servizi Sanitari, Trento, Italy

14 Azienda ULSS n 1 Dolomiti, Belluno, Italy

15 Independent Statistician, Solagna, Italy

Summary

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Prior appendectomy is associated with distinct immune microenvironment changes in rectal carcinoma tissue and adjacent normal mucosa, including altered expression of HLA-ABC, CTLA4, and CD86.

In a cohort of 163 rectal cancer patients (41 with prior appendectomy), normal mucosa from appendectomy patients showed higher epithelial HLA-abc expression (p=0.04) and higher CTLA4 expression on CD3+ T cells (p=0.03) compared to appendectomy-naive patients.
Within rectal cancer tissue, patients with prior appendectomy had higher CD86 expression on epithelial cancer cells (p=0.03).
In therapy-naive patients, prior appendectomy was associated with higher CD4+ T helper activation (CD25 expression, p=0.04), with trends toward higher epithelial HLA-abc (p=0.06) and T cell CTLA-4 (p=0.08).
In patients who received neoadjuvant therapy, prior appendectomy was associated with lower PD-L1 levels on leukocyte surfaces (p=0.039).
The authors interpret these findings as supporting the hypothesis that appendectomy may alter epithelial-intraepithelial T cell interactions in rectal cancer microenvironments.

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Abstract

Introduction

Although traditionally regarded as a vestigial organ, the appendix is increasingly recognized for its potential role in modulating gut immunity and contributing to the development of significant inflammatory or neoplastic intestinal diseases [1]. However, the underlying mechanisms linking the appendix to these immune and pathological processes remain poorly understood. This study aimed to investigate the relationship between the local immune microenvironment and the mutational landscape of rectal carcinomas in the context of prior or progressive appendectomy status.

Aims & Methods

This study aimed to investigate the relationship between the local immune microenvironment and the mutational landscape of rectal carcinomas in the context of prior or progressive appendectomy status.
Patients enrolled in the IMMUNOREACT 1 and 2 studies (NCT04915326 and NCT04917263, respectively) were stratified according to appendectomy status. Immunohistochemical analysis was performed to assess a range of immune markers, including CD3, CD4, CD8, CD8β, Tbet, FoxP3, PD-L1, MSH6, PMS2, and CD80. Additionally, flow cytometry was employed to evaluate the percentage of epithelial cells expressing CD80, CD86, CD40, HLA-ABC, or HLA-DR, as well as to quantify activated CD8+ T cells, CD4+ Th1 cells, and regulatory T cells (Tregs). Non-parametric tests were used.

Results

In this cohort, 41 patients out of 163 underwent appendectomy for other causes (e.g. appendicitis) before having been diagnosed with a rectal cancer. When the two groups were compared for microenvironmental features, in the normal mucosa of patients who had a previous appendectomy there was a higher rate of epithelial cells expressing HLA-abc (p=0,04) and a higher expression of CTLA4 on CD3+ T cells (p=0,03) when compared to appendectomy-naive patients. Within the rectal cancer tissue, a higher expression of CD86 on epithelial (cancer) cells expression was found in patients who already had an appendectomy (p=0,03). In therapy naïve patients, previous appendectomy was associated to a higher activation of CD4+ T helpers (expression of CD25), (p=0.04), and to tendency of higher rate of epithelial cells expressing HLA-abc (p=0,06) and of T cells expressing CTLA-4 (p=0.08). In patients who had neoadjuvant therapy, lower level of PD-L1 on the leukocyte surface were observed in patients who had a previous appendectomy (p=0.039).

Conclusion

Our findings suggest that prior appendectomy may be associated with distinct alterations in the local immune microenvironment of rectal carcinomas, including increased expression of antigen-presenting and immune-regulatory molecules such as HLA-BC, CTLA4, and CD86. In therapy naïve patients, appendectomy seem to increase the activation of T helper cells counterbalanced by an increase of the number of T cells expressing CTLA-4. These immune shifts in both tumour tissue and adjacent healthy mucosa support the hypothesis that the appendectomy may in somehow alter the interaction between epithelia cells and intraepithelial T cells. Further trancriptomic analysis will shed light on the effect of appendectomy on mucosal microevironment of rectal cancer patients.

References

1. Collard MK, Tourneur-Marsille J, Uzzan M, Albuquerque M, Roy M, Dumay A, Freund JN, Hugot JP, Guedj N, Treton X, Panis Y, Ogier-Denis E. The Appendix Orchestrates T-Cell Mediated Immunosurveillance in Colitis-Associated Cancer. Cell Mol Gastroenterol Hepatol. 2023;15(3):665-687.

Conference

UEG Week Berlin 2025

Topics

Colorectal

Submission format

Abstract

Session

Colorectal cancer: Working around genetics and complications (Posters)

Citation

United European Gastroenterology Journal 2025; 13 (Supplement 8)

Published

2025

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Unraveling Steatotic Liver Diseases: Insights from Amino Acid and TCA Cycle Metabolomics

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Unraveling Steatotic Liver Diseases: Insights from Amino Acid and TCA Cycle Metabolomics

Masaaki Mino 1

Affiliations

1 Kohnodai Hospital, National Center for Global Health and Medicine, Kōnodai, Japan

Summary

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Abstract

Event

UEG Week Berlin 2025

Topics

Hepatobiliary

Session

Rising Stars from Europe and Japan: Metabolic liver diseases

Citation

United European Gastroenterology Journal 2025; 13 (Supplement 8)

Published

2025

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József Maléth

Laudatio UEG Lifetime Achievement Awardee

Mario Dinis-Ribeiro

Accreditation for a newly recruited endoscopy nurse

Anne Vidil

UEG Presentation

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Time management: How can we save time?

Deepa Jegadish Gandhi 1

Affiliations

1 Mater Misericordiae University Hospital, Dublin, Ireland

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Abstract

Event

UEG Week Berlin 2025

Topics

Nurses

Session

Staff safety and management

Citation

United European Gastroenterology Journal 2025; 13 (Supplement 8)

Published

2025

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József Maléth

Laudatio UEG Lifetime Achievement Awardee

Mario Dinis-Ribeiro

Accreditation for a newly recruited endoscopy nurse

Anne Vidil

UEG Presentation

Share via Email Share on Facebook Share on X Share on LinkedIn Share on Bluesky

European survey on physical and mental health in endoscopy

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European survey on physical and mental health in endoscopy

Ute Pfeifer 1

Affiliations

1 Evangel. Krankenhaus Düsseldorf Abt. Endoskopie, Düsseldorf, Germany

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Abstract

Event

UEG Week Berlin 2025

Topics

Nurses

Session

Staff safety and management

Citation

United European Gastroenterology Journal 2025; 13 (Supplement 8)

Published

2025

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Laudatio UEG Lifetime Achievement Awardee

Mario Dinis-Ribeiro

Accreditation for a newly recruited endoscopy nurse

Anne Vidil

UEG Presentation

Share via Email Share on Facebook Share on X Share on LinkedIn Share on Bluesky

STORE-INDEPENDENT ORAI1 SIGNALING DEFINES BASAL CFTR FUNCTION IN SECRETORY EPITHELIA

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STORE-INDEPENDENT ORAI1 SIGNALING DEFINES BASAL CFTR FUNCTION IN SECRETORY EPITHELIA

Aletta Kata Kiss 1, Arpad Varga 1, Marietta Görög 2, Tamara Madacsy 1, Woo Young Chung 3, Petra Pallagi 1, Viktória Szabó 1, Petra Susánszki 2, Enikő Kúthy-Sutus 1, Szintia Barnai 2, Dániel Varga 4, Péter Bíró 4, Ingrid Hegnes Sendstad 1, Tim Crul 1, Boldizsár Jójárt 1, Bálint Tél 5, Anita Balazs 6, György Lázár 4, Miklós Erdélyi 4, Shmuel Muallem 3, József Maléth 1

Affiliations

1 University of Szeged, Szeged, Hungary|||University of Szeged, Szeged, Hungary|||University of Szeged, Szeged, Hungary

2 University of Szeged, Szeged, Hungary|||University of Szeged, Szeged, Hungary

3 National Institutes of Health, Bethesda, United States

4 University of Szeged, Szeged, Hungary

5 University of Szeged, Szeged, Hungary|||University of Szeged, Szeged, Hungary|||Semmelweis University, Budapest, Hungary

6 Charité Universitätsmedizin Berlin, Berlin, Germany

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Abstract

Introduction

Cystic fibrosis transmembrane conductance regulator (CFTR) is essential for epithelial ion and fluid secretion. While CFTR is primarily activated via cAMP/PKA signaling, the role of intracellular Ca²⁺ signaling—particularly through store-independent mechanisms—remains incompletely understood.

Aims & Methods

We aimed to elucidate how store-independent Ca²⁺ influx via Orai1 regulates basal CFTR activity in primary polarized epithelial cells.
To investigate this we used mouse and human pancreatic, hepatic and airway organoids, we combined RNA-sequencing, immunofluorescence, Fura-2 and MQAE dye-based Ca²⁺/Cl⁻ imaging, FLIM-FRET, dSTORM microscopy, and in vivo fluid secretion assays. Genetic silencing and pharmacological inhibition (CM5480) were employed to dissect Orai1-dependent signaling pathways.

Results

We discovered that Orai1 mediates a constitutive, store-independent Ca²⁺ influx sustained by SPCA2. This influx occurs independently of ER Ca²⁺ depletion but requires STIM1. Super-resolution imaging revealed Orai1 clustering into discrete apical membrane nanodomains that colocalize with SPCA2, STIM1, PKA, and CFTR. Functional assays confirmed that this localized signaling complex maintains basal CFTR activity. Inhibiting Orai1 reduced basal Cl⁻ and HCO₃⁻ secretion, disrupted mitochondrial ATP production, and suppressed pancreatic ductal fluid secretion in vitro and in vivo. Further, the stimulatory effect of this Ca²⁺ influx on CFTR was mediated via Ca²⁺/calmodulin-sensitive adenylyl cyclases (AC1, 3, 8), which were co-clustered with Orai1 and CFTR. Their knockdown phenocopied Orai1 inhibition, and no additive effect was observed when both pathways were blocked.

Conclusion

Our results reveal a previously unrecognized SPCA2–STIM1–Orai1 nanodomain that drives store-independent Ca²⁺ influx, activating CFTR via local cAMP production in the absence of neurohormonal stimuli. This regulatory module is conserved across secretory epithelia and represents a key mechanism for maintaining basal fluid secretion and epithelial homeostasis.

Event

UEG Week Berlin 2025

Topics

Mechanisms & Personalised Medicine Pancreas

Submission format

Abstract

Session

Cogulation, lipids and bugs: All relevant for pancreatitis

Citation

United European Gastroenterology Journal 2025; 13 (Supplement 8)

Published

2025

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Laudatio UEG Lifetime Achievement Awardee

This content is part of Gutflix. Log in with your myUEG account, or create one free, to watch it.

Not sure what you can access? Learn more about account types.

Laudatio UEG Lifetime Achievement Awardee

Mario Dinis-Ribeiro 1

Affiliations

1 IPO Porto, Porto, Portugal

Summary

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Abstract

Event

UEG Week Berlin 2025

Topics

Immunology

Session

Opening Plenary

Citation

United European Gastroenterology Journal 2025; 13 (Supplement 8)

Published

2025

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Unraveling Steatotic Liver Diseases: Insights from Amino Acid and TCA Cycle Metabolomics

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European survey on physical and mental health in endoscopy

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STORE-INDEPENDENT ORAI1 SIGNALING DEFINES BASAL CFTR FUNCTION IN SECRETORY EPITHELIA

József Maléth

Laudatio UEG Lifetime Achievement Awardee

Mario Dinis-Ribeiro

Accreditation for a newly recruited endoscopy nurse

Anne Vidil

UEG Presentation

Share via Email Share on Facebook Share on X Share on LinkedIn Share on Bluesky

Accreditation for a newly recruited endoscopy nurse

This content is part of Gutflix. Log in with your myUEG account, or create one free, to watch it.

Not sure what you can access? Learn more about account types.

Accreditation for a newly recruited endoscopy nurse

Anne Vidil 1

Affiliations

1 None, None, France

Summary

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Abstract

Event

UEG Week Berlin 2025

Topics

Nurses

Session

Education: Important tool for quality in endoscopy

Citation

United European Gastroenterology Journal 2025; 13 (Supplement 8)

Published

2025

More Like This:

Unraveling Steatotic Liver Diseases: Insights from Amino Acid and TCA Cycle Metabolomics

Masaaki Mino

Time management: How can we save time?

Deepa Jegadish Gandhi

European survey on physical and mental health in endoscopy

Ute Pfeifer

STORE-INDEPENDENT ORAI1 SIGNALING DEFINES BASAL CFTR FUNCTION IN SECRETORY EPITHELIA

József Maléth

Laudatio UEG Lifetime Achievement Awardee

Mario Dinis-Ribeiro

Accreditation for a newly recruited endoscopy nurse

Anne Vidil