Topical Application of Temperature-Sensitive Gel Containing Caerin 1.1 and 1.9 Peptides on TC-1 Tumour-Bearing Mice Induced High-Level Immune Response in the Tumour Microenvironment

Guoying Ni,Ming Q Wei,Conor E Fogarty,Pingping Zhang,Ping Zhang,Ying Liu,Hejie Li,Tianfang Wang,Shu Chen,Xiaosong Liu,Xiaolian Wu,Guoqiang Chen

doi:10.3389/fonc.2021.754770

Abstract

The development of topical cream drugs that increase the immune activation of tumour-infiltrating lymphocytes against tumour and chronic viral infection-associated lesions is of great immunotherapeutic significance. This study demonstrates that the topical application of a temperature-sensitive gel containing caerin 1.1 and 1.9 peptides reduces nearly 50% of the tumour weight of HPV16 E6/E7-transformed TC-1 tumour-bearing mice via improving the tumour microenvironment. Confocal microscopy confirms the time-dependent penetration of caerin 1.9 through the epidermal layer of the ear skin structure of mice. Single-cell transcriptomic analysis shows that the caerin 1.1/1.9 gel expands the populations with high immune activation level and largely stimulates the pro-inflammatory activity of NK and dendritic cells. Closely associated with INFα response, Cebpb seems to play a key role in altering the function of all Arg1hi macrophages in the caerin group. In addition, the caerin gel treatment recruits almost two-fold more activated CD8+ T cells to the TME, relative to the untreated tumour, which shows a synergistic effect derived from the regulation of S1pr1, Ccr7, Ms4a4b and Gimap family expression. The TMT10plex-labelling proteomic quantification further demonstrates the activation of interferon-alpha/beta secretion and response to cytokine stimulus by the caerin gel, while the protein contents of several key regulators were elevated by more than 30%, such as Cd5l, Gzma, Ifit1, Irf9 and Stat1. Computational integration of the proteome with the single-cell transcriptome consistently suggested greater activation of NK and T cells with the topical application of caerin peptide gel.

Highlights

Among 14 million new cancer cases reported worldwide in 2012, human papillomavirus (HPV) infection-associated cancers accounted for 4.6% of total cancers and nearly 30% of infection-related cancers [1]
High-risk HPV infection is related to a fraction of head-and-neck epithelial carcinoma in both developed and undeveloped countries [2], whereas the linking between HPV with cancers of the anus, vulva, vagina and penis is evident
Caerin 1.1 and 1.9 were able to inhibit multiple types of tumour growth in vitro, resulting in the apoptosis of the tumour cells. They inhibited TC-1 tumour growth in vivo when locally injected to tumour, and the tumour inhibition effect is dependent on the existence of intact adaptive immune systems, as their tumour inhibition effect disappeared in Rag-/- mice, which lacks B and T cells [62] and mice depleted of T cells

Summary

Introduction

Among 14 million new cancer cases reported worldwide in 2012, human papillomavirus (HPV) infection-associated cancers accounted for 4.6% of total cancers and nearly 30% of infection-related cancers [1]. Genital warts (condyloma acuminate, venereal warts and anogenital warts) are one of the most common sexually transmitted diseases (STDs) resulting from infection with low-risk HPV, especially HPV6 and 11, which lead to approximately 90% of the cases [3,4,5]. There are significant side effects associated with the topical application of imiquimod, such as erythema, scabbing, itching and burning [13]. Imiquimod-associated adverse effects at non-application sites were reported, such as fever, vertigo or myalgia, as well as distant inflammatory mucosal reactions [14]. Alternative treatments with minimal side effects have been under investigation

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Results

Conclusion