Interleukin-6 Induced “Acute” Phenotypic Microenvironment Promotes Th1 Anti-Tumor Immunity in Cryo-Thermal Therapy Revealed By Shotgun and Parallel Reaction Monitoring Proteomics

Abstract

Cryo-thermal therapy has been emerged as a promising novel therapeutic strategy for advanced breast cancer, triggering higher incidence of tumor regression and enhanced remission of metastasis than routine treatments. To better understand its anti-tumor mechanism, we utilized a spontaneous metastatic mouse model and quantitative proteomics to compare N-glycoproteome changes in 94 serum samples with and without treatment. We quantified 231 highly confident N-glycosylated proteins using iTRAQ shotgun proteomics. Among them, 53 showed significantly discriminated regulatory patterns over the time course, in which the acute phase response emerged as the most enhanced pathway. The anti-tumor feature of the acute response was further investigated using parallel reaction monitoring target proteomics and flow cytometry on 23 of the 53 significant proteins. We found that cryo-thermal therapy reset the tumor chronic inflammation to an “acute” phenotype, with up-regulation of acute phase proteins including IL-6 as a key regulator. The IL-6 mediated “acute” phenotype transformed IL-4 and Treg-promoting ICOSL expression to Th1-promoting IFN-γ and IL-12 production, augmented complement system activation and CD86+MHCII+ dendritic cells maturation and enhanced the proliferation of Th1 memory cells. In addition, we found an increased production of tumor progression and metastatic inhibitory proteins under such “acute” environment, favoring the anti-metastatic effect. Moreover, cryo-thermal on tumors induced the strongest “acute” response compared to cryo/hyperthermia alone or cryo-thermal on healthy tissues, accompanying by the most pronounced anti-tumor immunological effect. In summary, we demonstrated that cryo-thermal therapy induced, IL-6 mediated “acute” microenvironment shifted the tumor chronic microenvironment from Th2 immunosuppressive and pro-tumorigenic to Th1 immunostimulatory and tumoricidal state. Moreover, the magnitude of “acute” and “danger” signals play a key role in determining the efficacy of anti-tumor activity.

The cryo-thermal therapy-induced IL-6-rich acute pro-inflammatory response promoted DCs phenotypic maturation as the prerequisite to CD4+T cell differentiation

“…In our previous study, a novel tumour therapeutic modality was developed for cryo-thermal therapy by alternating liquid nitrogen (LN2) cooling and radio frequency (RF) heating of tumour tissues [22][23][24][25][26][27]. Using a highly malignant murine 4T1 breast cancer xenograft model, we found that cryo-thermal therapy remodelled the tumour microenvironment from chronic inflammatory to an acute phenotype.…”

“…Using a highly malignant murine 4T1 breast cancer xenograft model, we found that cryo-thermal therapy remodelled the tumour microenvironment from chronic inflammatory to an acute phenotype. Furthermore, IL-6 was significantly up-regulated during the acute phase after the cryo-thermal therapy [27]. However, whether acute IL-6 induced is essential to the enhancement of anti-tumour immunity is still unclear.…”

Interleukin‐6/STAT3 signaling as a promising target to improve the efficacy of cancer immunotherapy

“…A previous paper demonstrated that IL‐6 produced by nonhematopoietic stromal cells acted cooperatively with soluble IL‐6Rα and thermally induced gp130 to promote E/P‐selectin‐dependent and ICAM‐1‐dependent extravasation of cytotoxic T cells in tumors and apoptosis of tumor targets . Another paper revealed that cryo‐thermal therapy induced IL‐6 at acute phase and shifted the tumor chronic microenvironment from Th2 immunosuppressive and pro‐tumorigenic to a Th1 immunostimulatory and tumoricidal state …”

“…(21) Another paper revealed that cryo-thermal therapy induced IL-6 at acute phase and shifted the tumor chronic microenvironment from Th2 immunosuppressive and pro-tumorigenic to a Th1 immunostimulatory and tumoricidal state. (22) Because the precise effects of IL-6 on human cancer immunology are unclear compared with those in mice, we have recently focused on the IL-6/STAT3-signaling pathway in human DC. (23) Our aim was to translate the information obtained by murine studies for clinical benefit in human subjects, and to demonstrate the possibility of improving cancer immunotherapy by regulation of the IL-6/STAT3 signaling pathway.…”

Oxidative stress induced autophagy in cancer associated fibroblast enhances proliferation and metabolism of colorectal cancer cells

“…Tumorigenesis was closely related with its microenvironment, which was identified as "cancer cell nests" constituted by malignant cancer cells, immune cells, stromal cells and their secreted cytokines. [7][8][9] Cancer associated fibroblasts (CAFs) of tumor microenvironment were reported to promote tumor initiation, progression and metastasis by interfering cellular metabolism. [10][11][12] CAFs could supply energy-rich metabolites such as lactate through aerobic glycolysis pathway, which can then be used by adjacent cancer cells for ATP generation and for biosynthesis.…”