title: 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.
authors.affiliation: 1. Key Laboratory of Systems Biomedicine (MOE), Shanghai Center for Systems Biomedicine, the School of Biomedical Engineering and MED-X Research Institute, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Xue
authors.affiliation: 1. Key Laboratory of Systems Biomedicine (MOE), Shanghai Center for Systems Biomedicine, the School of Biomedical Engineering and MED-X Research Institute, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Liu
authors.affiliation: 2. Institute for Systems Biology, Seattle, WA, USA.
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: 1. Key Laboratory of Systems Biomedicine (MOE), Shanghai Center for Systems Biomedicine, the School of Biomedical Engineering and MED-X Research Institute, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Kun
authors.initials: K
authors.lastname: Liu
authors.affiliation: 2. Institute for Systems Biology, Seattle, WA, USA.
authors.forename: Li
authors.initials: L
authors.lastname: Yang
authors.affiliation: 2. Institute for Systems Biology, Seattle, WA, USA.
authors.forename: Robert L
authors.initials: RL
authors.lastname: Moritz
authors.affiliation: 1. Key Laboratory of Systems Biomedicine (MOE), Shanghai Center for Systems Biomedicine, the School of Biomedical Engineering and MED-X Research Institute, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Yan
authors.affiliation: 1. Key Laboratory of Systems Biomedicine (MOE), Shanghai Center for Systems Biomedicine, the School of Biomedical Engineering and MED-X Research Institute, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Lisa X
authors.initials: LX
authors.lastname: Xu
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D017679:Cryotherapy; D004195:Disease Models, Animal; D006979:Hyperthermia, Induced; D015850:Interleukin-6 / Q000378:metabolism*; D051379:Mice; D020543:Proteome / Q000032:analysis*; D018417:Th1 Cells / Q000276:immunology*; D059016:Tumor Microenvironment
chemical_list: D015850:Interleukin-6; D020543:Proteome; C577686:interleukin-6, mouse
keywords: Th1 anti-tumor immunity.; cryo-thermal therapy; interleukin-6; parallel reaction monitoring; “acute” phase response