Altering inflammatory states of immune cells in vivo by modulating cellular activation states
An activated state, immune cell technology, applied in chemical instruments and methods, carrier binding/immobilizing peptides, biochemical equipment and methods, etc., can solve problems such as dangerous side effects and insufficient penetration of tumor microenvironment by therapy
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Embodiment 1
[0329] Example 1. Materials and methods. PbAE synthesis. A method for synthesizing the polymer was previously described (Mangraviti A et al. (2015) ACS Nano 9:1236-1249). Mix 1,4-butanediol diacrylate with 4-amino-1-butanol in a 1:1 molar ratio of diacrylate to amine monomer. Acrylate-terminated poly(4-amino-1-butanol-co-1,4-butanediol diacrylate) was formed by heating the mixture to 90° C. for 24 hours with stirring. 2.3 g of this polymer were dissolved in 2 mL tetrahydrofuran (THF). To form the piperazine-terminated 447 polymer, 786 mg 1-(3-aminopropyl)-4-methylpiperazine in 13 mL THF was added to the polymer / THF solution and incubated at room temperature (RT) Stir for 2 hours. The capped polymer was precipitated with 5 volumes of diethyl ether, washed with 2 volumes of fresh ether, and dried under vacuum for 1 day. Neat polymer was dissolved in dimethyl sulfoxide (DMSO) to a concentration of 100 mg / mL and stored at -20 °C.
[0330] PGA is conjugated to di-mannose. α-...
Embodiment 2
[0354] Example 2. The therapeutic effect of NP-delivered M1-promoting gene on disseminated ovarian cancer. To evaluate this therapeutic approach in a clinically relevant in vivo testing system, a model recapitulating advanced, unresectable ovarian tumors in C57BL / 6 mice was used; these animals were injected with ID8 ovarian cancer cells, which Cells were labeled with luciferase to enable serial bioluminescence imaging of tumor growth (Liao JB et al (2015) J Immunother Cancer 3:16; Stephan SB et al (2015) Nat Biotechnol 33:97-101). Tumors were allowed to establish for two weeks. By this stage, mice had developed nodules throughout the peritoneal wall and mesentery. Animals were divided into 3 groups, which received PBS (control), GFPNP (sham), or IRF5 / IKKβNP treatment at an intraperitoneal dose of 100 μg mRNA / mouse / week for 9 weeks ( Figure 4A ). Disease regression was observed in the IRF5 / IKKβNP treated group and was eventually cleared in 40% of the animals (median surv...
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