Relaxin-encapsulated polymeric metformin nanoparticles remodel tumor immune microenvironment by reducing CAFs for efficient triple-negative breast cancer immunotherapy

Cancer-associated fibroblasts(CAFs)are one of the most abundant stromal cells in the tumor microenvironment which mediate desmoplastic response and are the primary driver for an immunosuppressive microenvironment,leading to the failure of triple-negative breast cancer(TNBC)immunotherapy.Therefore,depleting CAFs may enhance the effect of immunotherapy(such as PD-L1 antibody).Relaxin(RLN)has been demonstrated to significantly improve transforming growth factor-β(TGF-β)induced CAFs activation and tumor immunosuppressive microenvironment.However,the short half-life and systemic vasodilation of RLN limit its in vivo efficacy.Here,plasmid encoding relaxin(pRLN)to locally express RLN was delivered with a new positively charged polymer named polymeric metformin(PolyMet),which could increase gene transfer efficiency significantly and have low toxicity that have been certified by our lab before.In order to improve the stability of pRLN in vivo,this complex was further formed lipid poly-γ-glutamic acid(PGA)/PolyMetpRLN nanoparticle(LPPR).The particle size of LPPR was 205.5±2.9 nm,and the zeta potential was+55.4±1.6 mV.LPPR displayed excellent tumor penetrating efficacy and weaken proliferation of CAFs in 4T1luc/CAFs tumor spheres in vitro.In vivo,it could reverse aberrantly activated CAFs by decreasing the expression of profibrogenic cytokine and remove the physical barrier to reshape the tumor stromal microenvironment,which enabled a 2.2-fold increase in cytotoxic T cell infiltration within the tumor and a decrease in immunosuppressive cells infiltration.Thus,LPPR was observed retarded tumor growth by itself in the 4T1 tumor bearing-mouse,and the reshaped immune microenvironment further led to facilitate antitumor effect when it combined with PD-L1 antibody(aPD-L1).Altogether,this study presented a novel therapeutic approach against tumor stroma using LPPR to achieve a combination regimen with immune checkpoint blockade therapy against the desmoplastic TNBC model.

Calculation example of full cycle carbon emission of super deep foundation pit and carbon reduction measures

The determination of carbon emission from foundation pit engineering is a tough and complex project owing to its characteristics including large material consumption,short use time,difficult recycling and no operation stage.To overcome these limitations,the calculation boundary and calculation method for carbon emission of foundation pit project are defined in this paper,which is successfully applied in the carbon emission analysis of the actual engineering project,i.e.the construction of large-scale foundation pit of Kunming comprehensive transportation international hub.All the carbon emissions coresponding to four working stages including building materials production,building materials transportation,construction and demolition were calculated and anatomized.The results revealed that the content of CO_(2) released in the stage of building materials production accounts for 89.3%of the total carbon emission,which means the amount of building materials consumed in the engineering project is the crucial factor to control the carbon emission.Besides,two kinds of carbon reduction measures,i.e.optimization design of support scheme and recycling waste materials of internal support demolition,were explored by analyzing the proportion and average value of carbon emission from different sub project of the support structure.A pronounced effect of carbon reduction was achieved.Furthermore,both a fast calculation method of carbon emission factor of unit work volume and general carbon reduction measures are proposed in this paper,which could provide a reference and new viewpoint for the engineers and designers to calculate and analyze the carbon emission and to take effective carbon reduction measures.