Glycocalyx-Mimicking Nanoparticles Improve Anti-PD-L1 Cancer Immunotherapy through Reversion of Tumor-Associated Macrophages

Immune checkpoint blockade by anti-PD-L1 monoclonal antibody (αPD-L1) has achieved unprecedented clinical benefits in certain cancers, whereas the therapeutic efficacy is often hindered by immunosuppressive tumor microenvironment mediated by tumor-associated macrophages (TAMs), which leads to innate resistance to this approach. To improve checkpoint blockade efficacy, the amphiphilic diblock copolymers poly­(mannopyranoside/galactopyranoside methacrylate)-<i>block</i>-polystyrene are prepared by RAFT polymerization, which are sequentially self-assembled into glycocalyx-mimicking nanoparticles (GNPs) to neutralize TAMs. It is shown that GNPs can be specifically internalized by TAMs via lectin receptors, which results in upregulation of immunostimulatory IL-12 and downregulation of immunosuppressive IL-10, arginase 1, and CCL22, indicating functional reversion of protumor TAMs toward antitumor phenotype. The reversion of TAMs is proved to be mainly controlled by suppressing STAT6 and activating NF-κB phosphorylation. In vivo therapeutic studies have demonstrated that GNPs significantly enhance the therapeutic efficacy of αPD-L1 cancer therapy by reduction of tumor burden. Moreover, combination therapies with GNPs and αPD-L1 greatly improve immunosuppressive tumor microenvironment by reciprocal modulation of tumor-infiltrating effector and regulatory T cells. Notably, for the first time, our results demonstrate the reversion of TAMs and improvement of αPD-L1 cancer therapy by synthetic carbohydrate-containing nanomaterials. This research highlights a promising strategy for optimizing immune checkpoint blockade in cancer immunotherapy

Additional file 4: of Omics data reveal the unusual asexual-fruiting nature and secondary metabolic potentials of the medicinal fungus Cordyceps cicadae

Expression of putative mating and meiosis process-related genes by C. cicadae at different growth stages. (XLSX 13 kb

Effect of substrate concentrations on excretion rates (a), intracellular amounts (b), CL (c), and <i>f</i>_met (d) of Dio-7-G, Dio-3′-G, Chr-7-G, and Chr-4′-G.

<p>Three samples (500 μL) were obtained at 15, 30, and 60 min and replaced with fresh loading solution (500 μL) that contains diosmetin or chrysoeriol. The excretion rates of glucuronides were calculated from the slope of the amount-versus-time curves. The intracellular amounts of the glucuronides were determined at the end of the excretion experiments after the cells were washed twice with ice-cold HBSS. Each column corresponds to the average of three determinations with error bars representing the S.D. The “*” (for Dio-7-G or Chr-7-G) or “^#” (for Dio-3′-G or Chr-4′-G) symbol means a statistically significant difference between-group, at p < 0.05; “**” or “^##” means p < 0.01; “***” or “^###” means p < 0.001.</p

Effects of chemical inhibitors on diosmetin and metabolism and UGT1A9- and UGT1A1-mediated chrysoeriol metabolism in HLMs.

<p>Fig 4 A shows the inhibitory effects of phenylbutazone on the 7-O-glucuronide of diosmetin in HLMs and UGT1A6. Fig 4 B displays the inhibitory effects of carvacrol on the 3′-O-glucuronide of diosmetin in HLMs and UGT1A9. Fig 4 C presents the inhibitory effects of carvacrol on the 7-O-glucuronide of chrysoeriol in HLMs and UGT1A9. Fig 4 D shows the inhibitory effects of bilirubin on the 4′-O-glucuronides of chrysoeriol in HLMs and UGT1A1. Each column corresponds to the average of three determinations with error bars representing the S.D. The “*” symbol means a statistically significant difference compared with control at p < 0.05; “**” means p < 0.01; “***” means p < 0.001.</p

Kinetics of diosmetin and chrysoeriol glucuronidation by human expressed UGT enzymes.

<p>The curves are estimated on the basis of fitted parameters generated using the substrate inhibition (A, D) or Michaelis-Menten kinetics (B, C) in UGT1A6, UGT1A1, UGT1A9, and UGT1A9, respectively. The Eadie—Hofstee plots are shown in the right panal (a-d). Each data point corresponds to the average of three determinations with error bars representing the S.D.</p

Kinetic parameters of diosmetin and chrysoeriol glucuronidation by human expressed UGT enzymes.

<p>Kinetic parameters of diosmetin and chrysoeriol glucuronidation by human expressed UGT enzymes.</p

Basic information of patients with sporadic Parkinson’s disease (PD) and healthy controls.

<p>Basic information of patients with sporadic Parkinson’s disease (PD) and healthy controls.</p

Frequencies of HLA-DRB1 phenotypes and alleles in patients with Parkinson’s disease (PD) and healthy controls.

<p>Phenotype Frequency (Allele Frequency) was presented in every cell. “−”: the allele was not been detected. Phenotype Frequency: Percentage of individuals who have the allele (Individuals/N) in percentage format. Allele Frequency: Total number of copies of the allele in the population sample (Alleles/2N) in decimal format, Pc = correction of P value (Bonferroni adjustment), Pc<0.05 is considered as significant, ns = not significant. Patients had significant higher frequencies of HLA-DRB1*0301 and lower frequency of HLA-DRB1*0406 than healthy controls did.</p

The allele frequencies of HLA-DRB1*0301 and HLA-DRB1*0406 in populations from various ethnic regions.

<p>These data information were collected from the Allele Frequency Net Database <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048594#pone.0048594-GonzalezGalarza1" target="_blank">[52]</a> except reference 25,27–30,33–34,38–39,41,45,51. Allele Frequency: Total number of copies of the allele in the population sample (Alleles/2n) in decimal format. a: data from Chinese National Marrow Donor Program(CMDP), b: data from Tzu Chi Taiwan Marrow Donor Registry (TCTMDR), c: data from USA Colorado Univ. Cord Blood Bank, d: data from Poland DKMS, e: data from Umbilical Cord Blood Bank of Bacelona, f: data in Allele frequency net was calculated from Phenotype Frequencies assuming Hardy-Weinberg proportions.</p

Analysis of diosmetin, chrysoeriol, and their metabolites by UHPLC—MS/MS.

<p>Fig 1a and 1b show the chromatogram of the incubation samples of diosmetin and chrysoeriol by HLMs, respectively.</p