Colonic epithelial cathelicidin (LL-37) expression intensity is associated with progression of colorectal cancer and presence of CD8+ T cell infiltrate.

Manuel Salto-Tellez,Jacqueline A James,Mairi H Mclean,Stephanie G Craig,Graeme I Murray,Ji M Wang,Teizo Yoshimura,Abdo Alnabulsi,Ross J Porter,Matthew P Humphries

doi:10.1002/cjp2.222

Abstract

Colorectal cancer (CRC) remains a leading cause of cancer mortality. Here, we define the colonic epithelial expression of cathelicidin (LL‐37) in CRC. Cathelicidin exerts pleotropic effects including anti‐microbial and immunoregulatory functions. Genetic knockout of cathelicidin led to increased size and number of colorectal tumours in the azoxymethane‐induced murine model of CRC. We aimed to translate this to human disease. The expression of LL‐37 in a large (n = 650) fully characterised cohort of treatment‐naïve primary human colorectal tumours and 50 matched normal mucosa samples with associated clinical and pathological data (patient age, gender, tumour site, tumour stage [UICC], presence or absence of extra‐mural vascular invasion, tumour differentiation, mismatch repair protein status, and survival to 18 years) was assessed by immunohistochemistry. The biological consequences of LL‐37 expression on the epithelial barrier and immune cell phenotype were assessed using targeted quantitative PCR gene expression of epithelial permeability (CLDN2, CLDN4, OCLN, CDH1, and TJP1) and cytokine (IL‐1β, IL‐18, IL‐33, IL‐10, IL‐22, and IL‐27) genes in a human colon organoid model, and CD3+, CD4+, and CD8+ lymphocyte phenotyping by immunohistochemistry, respectively. Our data reveal that loss of cathelicidin is associated with human CRC progression, with a switch in expression intensity an early feature of CRC. LL‐37 expression intensity is associated with CD8+ T cell infiltrate, influenced by tumour characteristics including mismatch repair protein status. There was no effect on epithelial barrier gene expression. These data offer novel insights into the contribution of LL‐37 to the pathogenesis of CRC and as a therapeutic molecule.

Highlights

Human cathelicidin is an anti-microbial peptide, expressed by epithelial cells at mucosal surfaces and a wide array of immune cells [1]
This study aimed to define the epithelial cytoplasmic expression profile of cathelicidin linked to clinicopathological factors in human colorectal cancer (CRC), assess whether this could be a biomarker for prognosis, and explore the biological consequences of expression on the epithelial barrier and immune cell phenotype
We demonstrate the expression of LL-37 in a large (n = 650) fully characterised cohort of treatmentnaïve primary human colorectal tumours and 50 matched normal mucosa samples and show expression intensity of epithelial cathelicidin is associated with human CRC progression and CD8+ T cell infiltrate

Summary

Introduction

Human cathelicidin (and its bioactive derivative LL-37) is an anti-microbial peptide, expressed by epithelial cells at mucosal surfaces and a wide array of immune cells [1]. Genetic knockout of cathelicidin (CRAMP in the mouse) led to shorter crypt length implying a homeostatic role in colonic epithelial growth dynamics [5]. There is evidence to suggest a protective role of cathelicidin in colonic carcinogenesis through effects on growth dynamics, apoptosis and autophagy, angiogenesis, and fibroblast activity [1,2,8] and linked to the colonic microbiome [5]. Our previous data revealed that genetic knockout of CRAMP (murine cathelicidin) led to increased tumour size and number in the azoxymethane-induced murine model of colorectal cancer (CRC), suggesting a tumour-suppressive role [5]

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