Supplementary Materials

The PDF file includes:

  • Materials and Methods
  • Fig. S1. Biophysical homology screening using Molly font identifies conservation of 10-mer structural motif across HDPs, outer surface virulence factors, and collagens.
  • Fig. S2. Docking of biophysical 10-mer homology motifs onto C-type lectin receptors.
  • Fig. S3. Full-length CD206 model.
  • Fig. S4. RP-182 binds to CD206.
  • Fig. S5. RP-182 but not RP-426 induces a cellular thermostability shift of CD206 in human and murine M2 macrophages.
  • Fig. S6. Abbreviated synthesis scheme of RP-182 analogs with diazirine-containing phenylalanine and biotin.
  • Fig. S7. RP-182 binds to carbohydrate recognition domain 5 of the CD206 receptor.
  • Fig. S8. Gene expression changes induced by RP-182 in M1- and M2-polarized BMDMs.
  • Fig. S9. RNA-seq analysis of vehicle- versus RP-182-treated M2 macrophages and enrichment analysis of binding partners to CD206.
  • Fig. S10. RP-182 activates phagocytosis and phagolysosome formation in M2- but not M1-polarized human macrophages derived from PBMCs of healthy volunteers.
  • Fig. S11. RP-182 activates phagocytosis, autophagy, and apoptosis in BMDMs cocultured with conditioned medium from PANC1 cancer cells.
  • Fig. S12. RP-182 but not control peptide RP-426 induces phagocytosis in M2-polarized BMDMs.
  • Fig. S13. RP-182 reduces IKK-α and activates autophagy and caspase 8 in M2-polarized BMDMs.
  • Fig. S14. RP-182 activates autophagy and apoptosis in human M2 macrophages.
  • Fig. S15. RP-182 but not RP-426 induces cell death in M2-polarized macrophages.
  • Fig. S16. RP-182 does not inhibit growth of pluripotent progenitor cells, cancer cells, fibroblasts, or endothelial or dendritic cells.
  • Fig. S17. Gating strategy for the determination of CD86+ and CD206-positive CD11b+F4/80+Gr1 macrophage fractions.
  • Fig. S18. Gating strategy for the determination of M1 cytokine-positive CD11b+F/40+Gr1 macrophages and M1 and M2 marker expression profiles of M2 BMDMs isolated by FACS after treatment with RP-182.
  • Fig. S19. Gating strategy for the determination of cytokine- and checkpoint-positive cell fractions of CD86- and CD206-positive macrophage populations.
  • Fig. S20. M1 and M2 gene expression profiles of BMDMs isolated from B6.129P2-Mrc1tm1Mnz/J (CD206−/−) and wild-type C57BL/6 (CD206+/+) mice after polarization into M1 and M2.
  • Fig. S21. RP-182 induces binding of IQGAP1 to CD206 and recruitment of IQGAP1 to the cell membrane of M2-polarized macrophages.
  • Fig. S22. Blockade of RP-182–induced TNF signaling abrogates autocrine activation of apoptosis in M2-polarized macrophages.
  • Fig. S23. CD206 expression in human pancreatic cancer.
  • Fig. S24. Pretreatment of M2-polarized macrophages with RP-182 reduces induction of expression of EMT markers in KPC cancer cells upon coculture.
  • Fig. S25. Gating strategy for the identification of tumor-infiltrating CD8+ T cell and CD206high M2-like TAM fractions.
  • Fig. S26. RP-182 in combination with gemcitabine reduces CD206high monocytic MDSCs in autochthonous KPC tumors.
  • Fig. S27. Gating strategy for determination of PD-1, SIRP-α, and TNF-α–positive TAM populations in KPC mice.
  • Fig. S28. RP-182–induced CD86+ TAM populations in KPC mice have increased M1 cytokine and M1 marker expression.
  • Fig. S29. RP-182 induces phagocytosis, phagolysosome formation, and apoptosis in TAMs but not in CD11bCK19+ cancer cells.
  • Fig. S30. Tissue distribution and toxicity measures of RP-182 in mice.
  • Fig. S31. RP-182 induces cancer cell phagocytosis in CD86- but not CD206-positive M2 BMDMs.
  • Fig. S32. RP-182 restricts tumor growth of MDA-MB231 breast cancer and C4-2 prostate cancer xenografts grown in T cell–deficient mice.
  • Table S1. Origins of representative 10-mer biophysical homology peptide sequences within HDPs, virulence factors, and collagens.
  • Table S2. Relative binding affinities (in kcal/mol) of individual 10-mer homology sequence ligand-lectin receptor combinations by ClusPro web portal identifies CD206 as top binding target.
  • Table S3. List of primary and secondary antibodies used in immunocytochemistry, tissue immunohistochemistry, Western blots, immunoprecipitation, and flow cytometry experiments with manufacturers and catalog numbers.
  • Legends for movies S1 and S2
  • Legends for data files S1 to S3
  • Reference (46)

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Other Supplementary Material for this manuscript includes the following:

  • Movie S1 (.mov format). RP-182 increases phagocytosis of cancer cells.
  • Movie S2 (.mov format). Engulfment and internalization of cancer cells by RP-182–treated macrophages.
  • Data file S1 (.pdf format). Biophysical homology screening of AMPs and HDPs using Molly font.
  • Data file S2 (Microsoft Excel format). Characterization of binding partners of CD206 induced by RP-182 in M2 macrophages.
  • Data file S3 (Microsoft Excel format). Source data.