IL-7 pathway controls human T cell homing to the gut and culminates in inflammatory bowel disease mucosa

(1)

HAL Id: inserm-02161048

https://www.hal.inserm.fr/inserm-02161048

Submitted on 20 Jun 2019

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IL-7 pathway controls human T cell homing to the gut and culminates in inflammatory bowel disease mucosa

Lyssia Belarif, L. Kermarec, V. Daguin, Mary C., R. Danger, Kucik A., T. Macdonald, G. Blancho, P. Naveilhan, Bernard Vanhove, et al.

To cite this version:

Lyssia Belarif, L. Kermarec, V. Daguin, Mary C., R. Danger, et al.. IL-7 pathway controls human T cell homing to the gut and culminates in inflammatory bowel disease mucosa. FOCIS 2017, Jun 2017, Chicago, United States. �inserm-02161048�

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1

_{Ose Immunotherapeutics, Nantes, France;}

2

_{Centre de Recherche en Transplantation et Immunologie UMR 1064, INSERM, Université de Nantes, Nantes, France;}

3

_{Institut de Transplantation Urologie Néphrologie (ITUN), CHU Nantes, Nantes, France}

4

_{INSERM, UMR 913, Nantes F-44035, France;}

4

_{Blizard Institute, Barts and the London School of Medicine and Dentistry, London, UK.}

Conclusion

Interleukin-7 (IL-7) is a limiting and non-redundant non-classical cytokine produced essentially by

epithelial and stromal cells which regulates T-lymphocytes homeostasis. Almost all conventional mature T lymphocytes express the IL-7 receptor (IL-7R), with a particular exception for naturally-occurring regulatory T-cells (Tregs), constituting a rare opportunity to selectively target pathogenic effectors while preserving natural regulators. The signaling networks perpetuating chronic inflammatory bowel disease in man remain unclear. While in mice IL-7 is known to play a role in systemic inflammation, here we found that IL-7 specifically in humans controls α4/β7 integrin expression and imprints gut-homing specificity on T cells.

Introduction

L. Belarif

1,2,3

, L. Kermarec

4

, V. Daguin

1,2

, C. Mary

1,2,3

, R. Danger

1,2

, A. Kucik

5

, T. MacDonald

5

, G. Blancho

2,3

, P. Naveilhan

4

, B. Vanhove

1,2,3

, N. Poirier

1,2,3

IL-7 pathway controls human T cell homing to the gut

and culminates in inflammatory bowel disease mucosa

I- IL-7 controls human effector, but not regulatory T lymphocyte α4/β7 integrin expression

II- Anti-IL-7Ra delays colon inflammation in GVHD humanized mice model

_{III- Anti-IL-7Ra prevents acute colitis in humanized mice model}

IV- IL-7/IL-7R mRNA pathway is accumulated in colon biopsies from UC and CD patients and correlates with treatment-refractory disease

❖ GVHD was induced after infusion of 50.106

human PBMC in NSG mice. Anti-IL7Ra mAb significantly prolonged survival. Strikingly, colon inflammation and human T-cell infiltration in the colon were abolished, without impact on other target tissue of GVHD (intestine, liver and lung).

❖ Acute colitis was induced by intrarectal TNBS/Ethanol administration in humanized mice

reconstituted with human CD34+ _{cord blood}

hematopoietic stem cells. Anti-IL-7Ra mAb prevented colitis with an efficacy similar to Vedolizumab (anti- α4/β7 integrin mAb).

Results

1 3 5 7 10 0 200 400 600 800 1000 Days E x pr e s s ion ( M FI ) itga4-itgb7 10- 2 ₁₀- 1 ₁₀0 ₁₀1 0 20 40 60 80 100 120 aIL-7R (µg/ml) No rm a liz e d -e x p re s s io n ( % )

ITGA4/ITGB7

pSTAT5

itga4

itgb7

itga4

itgb7

CD3+CD4+ CD25-CD127High CD3+CD4+ CD25high_CD127Low It ga 4 e xp re ss io n (M FI ) It gb 7 e xp re ss io n (M FI ) It ga 4 e xp re ss io n (M FI ) It gb 7 e xp re ss io n (M FI ) Ex p re ss io n (M FI ) Ex p re ss io n (M FI ) Basal level

❖ ITGA4/ITGB7, two integrins implicated in the migration of T cells into the gut, are up-regulated by human recombinant IL-7 (rhIL-7) on effector (CD4+_CD25-_CD127high_{) but}

not on regulatory T cells (Tregs: CD4+_CD25high_CD127low_{) .}

❖ Up regulation of ITGA4/ITGB7 on effector (CD4+_CD25-_CD127high_{) T cells}

is blocked by anti-IL-7R antibodies. ❖ Induction of pSTA5 in response to IL-7

was observed in T cell and in Treg cells, demonstrating that in spite of low CD127 expression level, Tregs are able to respond to IL-7.

itga4 itgb7 medium 1 ng/ml 5 ng/ml 25 ng/ml 0 20 40 60 80 100 0 20 40 60 80 100 Time (Days) P er cen t su rvi val ctrl aIL-7R 0 1 2 3 H u m a n T c e ll i n fi ltra te s ctrl aIL-7R 7 8 9 10 Co lo n l e n g th (c m )

**

*

ctrl aIL-7R 0 5 10 15 H ito lo g ic a l S c o re

*

ctrl aIL-7R 0 1 2 3 H u m a n T c e ll i n fi lt ra te s Colon Intestine ctrl aIL-7R 0 1 2 3 H u m a n T c e ll i n fi ltra te s ctrl aIL-7R 0 1 2 3 H u m a n T c e ll i n fi ltra te s Liver Lung

*

Control aIL-7Rα

x10

IL-7R: 0.969 IL-7: 0.674 ITGA4: 0.840 ITGB7: 0.605 TNF: 0.640 TNFR1: 0.736 Active UC - Inflammed Active UC - healthy UC remission Non-IBD control

❖ IL-7/IL-7Ra pathway mRNA expression is significantly increased in the involved colon of CD and UC patients in comparaison with uninvolved biopsy from same patients or Non-IBD controls.

❖ IL-7Rα mRNA expression is correlated to ITGA4/ITGB7 mRNA expression in UC and CD patients : (ITGA4 : p< 0,0001 ; ITGA4 : p<0,0001 and ITGB7 : p<0,0001 ; ITGB7 : p<0,0001).

Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 10 20 30 40 m RN A e xp re s s io n (A .U.) UC non-responders (aTNF/IS) Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 2 4 6 8 m RN A e xp re s s io n (A .U.) UC non-responders (aTNF/IS) Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 1 2 3 4 5 m RN A e xp re s s io n (A .U.) UC non-responders (aTNF/IS) Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 20 40 60 80 100 m RN A e xp re s s io n (A .U.) CD non-responders (aTNF/IS) Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 5 10 15 m RN A e xp re s s io n (A .U.) CD non-responders (aTNF/IS) Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 1 2 3 4 m RN A e xp re s s io n (A .U.) UC non-responders (aTNF/IS) -4 -2 0 2 4 -4 -2 0 2 4 Log2 IL-7Ra L o g 2 I T G A 4 P < 0.0001 r = 0.70 P < 0.0001 r = 0.70 -4 -2 0 2 4 -2 -1 0 1 2 3 Log2 IL-7Ra L o g 2 I T G B 7 P < 0.0001 r = 0.66 P < 0.0001 r = 0.59

IL-7

IL-7Rα

γ-chain

*** *** *** * ** ** * * ** Crohn disease (CD) Ulcerative colitis (UC)

❖ 21 genes from the IL-7 signaling pathway were re-analyzed from a transcriptomic study of colon biopsies from UC patients. This IL-7 signature segregates into three distinct blocs according to the clinical status. Principal Component Analysis (PCA) of this IL-7 mucosal signature in UC patients displayed a clear and distinct separation between responders versus non-responder patients. IL-7Rα micro-array values discriminated UC patients with inflamed versus non-inflamed mucosa (ROC analysis: AUC = 96.9%).

❖ IL-7 controls α4/β7 integrin expression in human effector T lymphocytes, but not in Tregs

❖ Anti-IL7Ra mAb decreases specifically colon inflammation in humanized mice models

❖ Anti-IL7Ra mAb decreases IFNg production in human organ culture assay

❖ Transcripts of the IL-7 pathway accumulate in UC biopsies and correlate with absence of response to

treatments

Control α4β7 Ab IL7Rα Ab 0 1 2 3 4 5 6 7 8 9 10 0 1 2 3 Time (Days) Co li ti s s c o re 0 1 2 3 4 5 6 7 8 9 10 80 85 90 95 100 Time (Days)  W ei g h t (% ) p < 0.05 p < 0.05 p < 0.05 p < 0.05 human IL-7

V- Anti-IL7Ra mAb decreases ex-vivo

IFNg production by UC colon biopsies

❖ Colon biopsies from inflamed area of UC patients

(n=10) were cultured ex-vivo for 24 hours. These tissues spontaneously release proinflammatory cytokines such as IFNg (130± 19 pg/ml). Anti-IL7Rα mAb applied at 10 µg/ml in this organ culture assay significantly decreased inflammation as measured by IFNγ secretion level.

IL-7R FYN JAK3 STAT5B LCK ITGA4 PIK3CG IL2RG PTK2B ITGB7 CRLF2 LTBR TSLP EP300 NMI JAK1 IL-7 PI3K PIK3CA CREBBP BCL2 Group

a

b

Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 10 20 30 40 m RN A e xp re s s io n (A .U.) UC non-responders (aTNF/IS) Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 2 4 6 8 m RN A e xp re s s io n (A .U.) UC non-responders (aTNF/IS) Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 1 2 3 4 5 m RN A e xp re s s io n (A .U.) UC non-responders (aTNF/IS) Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 20 40 60 80 100 m RN A e xp re s s io n (A .U.) CD non-responders (aTNF/IS) Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 5 10 15 m RN A e xp re s s io n (A .U.) CD non-responders (aTNF/IS) Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 1 2 3 4 m RN A e xp re s s io n (A .U.) UC non-responders (aTNF/IS)

IL-7

IL-7Rα

γ-chain

IL-7

IL-7Rα

γ-chain

*** *** *** * ** ** * * **

a

b

Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 10 20 30 40 m RN A e xp re s s io n (A .U.) UC non-responders (aTNF/IS) Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 2 4 6 8 m RN A e xp re s s io n (A .U.) UC non-responders (aTNF/IS) Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 1 2 3 4 5 m RN A e xp re s s io n (A .U.) UC non-responders (aTNF/IS) Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 20 40 60 80 100 m RN A e xp re s s io n (A .U.) CD non-responders (aTNF/IS) Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 5 10 15 m RN A e xp re s s io n (A .U.) CD non-responders (aTNF/IS) Non -IBD co ntro ls Health y b iop sy Inflam med bio psy 0 1 2 3 4 m RN A e xp re s s io n (A .U.) UC non-responders (aTNF/IS)