EN
Project - B06:
The tolerogenic microenvironment in the liver facilitates metastasis formation
Project leader:
Anastasios Giannou, NP, MD, MSc, PhD
Summary
Metastasis is a common cause of cancer-related death. The majority of patients with colorectal cancer (CRC) suffer from metastases. Of note, liver metastasis formation is promoted by a tolerogenic micro-environment, which may further compromise immunotherapy efficacy. Here, we hypothesise that IL-10 is a key player in tolerance towards liver metastasis and may thus serve as an immunotherapeutic target in this context. This is based on our preliminary findings showing that Foxp3+ regulatory T cell (Treg)-derived IL-10 promotes CRC-derived liver metastasis (CRLM). Furthermore, we found that Foxp3+ Treg-derived IL-10 induces a subset of dysfunctional CD8+ T cells expressing TCF-1+. Based on this observation, we will initially focus on the mechanisms underlying the IL-10-mediated regulation of CD8+ T cells. In particular, we will decipher the role of PD-1 and TCF-1. To this end, we will use transgenic mice which allow the conditional, cell-specific deletion of the IL-10 receptor (IL-10Ra) in CD8+ T cells. CD8+ T cells will be characterized after liver metastasis induction using flow cytometry and functional in vitro assays. We will use bulk RNA sequencing in order to identify further mechanisms, which will be validated and then targeted using transgenic mice and in vitro assays. We specifically aim to examine whether IL-10 can be targeted to modulate liver tolerance and improve the immunotherapy response rate. Our long-term aim is to understand how liver tolerance can be broken to prevent or treat liver metastasis, while in parallel sustaining liver homeostasis and avoiding an enhanced risk for liver autoimmune and inflammatory conditions. Moreover, we will extend the understanding of IL-10-mediated CD8+ T-cell dysfunction in the context of liver metastasis receiving immunotherapy in mice and humans in the second funding period.
Project plan
Our central hypothesis is that IL-10 is a key player in tolerance towards liver metastasis and may thus serve as an immunotherapeutic target in this context. Our preliminary data indicate that Foxp3+ Treg-derived IL-10 promotes CRC-derived liver metastasis (CRLM). Additionally, Foxp3+ Treg-derived IL-10 could promote dysregulation of CD8+ T cells, with a skew towards TCF-1+ CD8+ T cells in the early phase during liver metastasis formation. On this basis, we will decipher the mechanisms underlying the IL-10-mediated regulation of dysfunctional CD8+ T cells. Our long-term aim is to examine whether IL-10 can be targeted therapeutically to modulate liver immune regulation aiming to improve immunotherapy response rates.
In order to test this hypothesis, our work programme has the following work packages (WP):
WP1: To investigate the mechanisms underlying the IL-10-mediated regulation of CD8+ T cells in liver metastasis formation.
WP2: To investigate how IL-10 mediated modulation of CD8+ T-cells impacts immunotherapy response in liver metastasis.
WP3: To examine whether IL-10 can serve as biomarker to break liver tolerance and improve immuno-therapy outcomes of CRC patients with liver metastasis.
Project related publications
Giannou AD#, Kempski J#, Shiri AM#, Lücke J, Zhang T, Zhao L, Zazara DE, Cortesi F, Riecken K, Amezcua Vesely MC, Low JS, Xu H, Kaffe E, Garcia-Perez L, Agalioti T, Yamada Y, Jungraithmayr W, Zigmond E, Karstens KF, Steglich B, Wagner J, Konczalla L, Carambia A, Schulze K, von Felden J, May P, Briukhovetska D, Bedke T, Brockmann L, Starzonek S, Lange T, Koch C, Riethdorf S, Pelczar P, Böttcher M, Sabihi M, Huber FJ, Reeh M, Grass JK, Wahib R, Seese H, Stüben BO, Fard-Aghaie M, Duprée A, Scognamiglio P, Plitzko G, Meiners J, Soukou S, Wittek A, Manthey C, Maroulis IC, Arck PC, Perez D, Gao B, Zarogiannis SG, Strowig T, Pasqualini R, Arap W, Gosálvez JS, Kobold S, Prinz I, Guse AH, Tachezy M, Ghadban T, Heumann A, Li J, Melling N, Mann O, Izbicki JR, Pantel K, Schumacher U, Lohse AW, Flavell RA, Gagliani N, Huber S. Tissue resident iNKT17 cells facilitate cancer cell extravasation in liver metastasis via interleukin-22. Immunity 2023;56:125-142.e12. doi: 10.1016/j.immuni.2022.12.014. Open access.
Briukhovetska D#, Suarez-Gosalvez J#, Voigt C#, Markota A, Giannou AD, Schübel M, Jobst J, Zhang T, Dörr J, Märkl F, Majed L, Müller PJ, May P, Gottschlich A, Tokarew N, Lücke J, Oner A, Schwerdtfeger M, Andreu-Sanz D, Grünmeier R, Seifert M, Michaelides S, Hristov M, König LM, Cadilha BL, Mikhaylov O, Anders HJ, Rothenfusser S, Flavell RA, Cerezo-Wallis D, Tejedo C, Soengas MS, Bald T, Huber S, Endres S, Kobold S. T cell-derived interleukin-22 drives the expression of CD155 by cancer cells to suppress NK cell function and promote metastasis. Immunity 2023;56:143-161.e11. doi: 10.1016/j.immuni.2022.12.010. Open access.
Kleinschmidt D, Giannou AD, McGee HM, Kempski J, Steglich B, Huber FJ, Ernst TM, Shiri AM, Wegscheid C, Tasika E, Hübener P, Huber P, Bedke T, Steffens N, Agalioti T, Fuchs T, Noll J, Lotter H, Tiegs G, Lohse AW, Axelrod JH, Galun E, Flavell RA, Gagliani N, Huber S. A Protective Function of IL-22BP in Ischemia Reperfusion and Acetaminophen-Induced Liver Injury. J Immunol 2017;199:4078–90. doi: 10.4049/jimmunol.1700587. Epub 2017 Nov 6.
Kempski J#, Giannou AD#, Riecken K, Zhao L, Steglich B, Lücke J, Garcia-Perez L, Karstens KF, Wöstemeier A, Nawrocki M, Pelczar P, Witkowski M, Nilsson S, Konczalla L, Shiri AM, Kempska J, Wahib R, Brockmann L, Huber P, Gnirck AC, Turner JE, Zazara DE, Arck PC, Stein A, Simon R, Daubmann A, Meiners J, Perez D, Strowig T, Koni P, Kruglov AA, Sauter G, Izbicki JR, Guse AH, Rösch T, Lohse AW, Flavell RA, Gagliani N#, Huber S#. IL22BP Mediates the Antitumour Effects of Lymphotoxin Against Colorectal Tumours in Mice and Humans. Gastroenterology 2020;159:1417-1430.e3. doi: 10.1053/j.gastro.2020.06.033. Epub 2020 Jun 22. Open access.
Zhao L#, Giannou AD#, Xu Y#, Shiri AM, Liebold I, Steglich B, Bedke T, Zhang T, Lücke J, Scognamiglio P, Kempski J, Woestemeier A, Chen J, Agalioti T, Zazara DE, Lindner D, Janning M, Hennigs JK, Jagirdar RM, Kotsiou OS, Zarogiannis SG, Kobayashi Y, Izbicki JR, Ghosh S, Rothlin CV, Bosurgi L#, Huber S#, Gagliani N#. Efferocytosis fuels malignant pleural effusion through TIMP1. Sci Adv 2021;7:eabd6734. doi: 10.1126/sciadv.abd6734.
Zhang T#, Wahib R#, Zazara D, Lücke J, Shiri A, Kempski J, Zhao L, Agalioti T, Machicote A, Giannou O, Belios I, Jia R, Zhang S, Tintelnot J, Seese H, Grass J, Mercanoglu B, Stern L, Scognamiglio P, Fard-Aghaie M, Seeger P, Wakker J, Kemper M, Brunswig B, Duprée A, Lykoudis PM , Pikouli A, Giorgakis E, Stringa P, Lausada N, Gentilini MV, Gondolesi GE, Bachmann K, Busch P, Grotelüschen R, Maroulis IC, Arck PC, Nakano R, Thomson AW, Ghadban T, Tachezy M, Melling N, Achilles E, Nickel F, Hackert T, Mann O, Izbicki JR, Li J, Gagliani N, Huber S#, Giannou AD#. CD4+ T cell-derived IL-22 enhances liver metastasis by promoting angiogenesis. OncoImmunology 2023;12. doi: 10.1080/2162402X.2023.2269634 .
Giannou AD#, Ohm B#, Zazara DE, Lücke J, Zhang T, Sabihi M, Seeger P, Oh J, Grotelüschen R, Busch P, Mann O, Hackert T, Izbicki JR, Yamada Y, Huber S#, Jungraithmayr W#. Protocol for orthotopic single-lung transplantation in mice as a tool for lung metastasis studies. STAR Protoc 2023;4:102701.doi: 10.1016/j.xpro.2023.102701. Epub 2023 Nov 10. Open access.
Lücke J#, Zhang T#, Zazara DE, Seeger P, Izbicki JR, Hackert T, Huber S#, Giannou AD#. Protocol for generating lung and liver metastasis in mice using models that bypass intravasation. STAR Protoc 2024;5:102696. doi: 10.1016/j.xpro.2023.102696. Epub 2024 Jan 18. Open access.
Lücke J#, Mercanoglu B#, Zhang T, Zazara DE, Zigmond E, Seeger P, Mann O, Izbicki JR, Hackert T, Huber S#, Giannou AD#. Mouse models of spontaneous liver and lung metastasis for colorectal cancer. STAR Protoc 2024;5:102811. doi: 10.1016/j.xpro.2023.102811. Epub 2024 Jan 17. Open access.
Shiri MA#, Zhang T#, Bedke T, Zazara DE, Zhao L, Lücke J, Sabihi M, Kempski J, Agalioti T, Mikołaj N, Xu Y, Riecken K, Liebold I, Brockmann L, Konczalla L, Bosurgi L, Mercanoglu B, Seeger P, Lykoudis PM, Arck PC, Fehse B, Busch P, Grotelüschen R, Mann O, Izbicki JR, Hackert T, Gagliani N, Giannou AD#, Huber S#. IL-10-mediated PD-L1 upregulation dampens antitumour immunity and promotes liver metastasis. J Hepatol. 2024 Apr;80(4):634-644. doi: 10.1016/j.jhep.2023.12.015. Epub 2023 Dec 30. Open access.
# equally contributing authors