Anni Lepland

Molecular tumor boards are supported by clinical decision support system (CDSS) tools for the interpretation of complex molecular data towards personalized treatments. Current CDSS tools are genomics-focused and characterized by tight couplings between data ingestion, guideline-based reasoning logic and curated knowledgebases. Next-generation CDSS solutions should integrate multimodal data and learn from outcomes in an interoperable ecosystem, leveraging explainable, trustworthy output from AI models through rigorous clinical validation processes.

Although many studies have explored the depletion of tumor-associated macrophages (TAM) as a therapeutic strategy for solid tumors, currently available compounds suffer from poor efficacy and dose-limiting side effects. Here, we developed a novel TAM-depleting agent (“OximUNO”) that specifically targets CD206+ TAMs and demonstrated efficacy in a triple-negative breast cancer (TNBC) mouse model. OximUNO comprises a star-shaped polyglutamate (St-PGA) decorated with the CD206-targeting peptide mUNO that carries the chemotherapeutic drug doxorubicin (DOX). In the TNBC model, a fluorescently labeled mUNO-decorated St-PGA homed to CD206+ TAMs within primary lesions and metastases. OximUNO exhibited no acute liver or kidney toxicity in vivo. Treatment with OximUNO reduced the progression of primary tumor lesions and pulmonary metastases, significantly diminished the number of CD206+ TAMs and increased the CD8/FOXP3 expression ratio (indicating immunomodulation). Our findings suggest the potential benefit of OximUNO as a TAM-depleting agent for TNBC treatment. Importantly, our studies also represent a novel design of a peptide-targeted St-PGA as a targeted therapeutic nanoconjugate.Significance:A peptide-targeted nanoformulation of DOX exclusively eliminates mannose receptor+ TAMs in breast cancer models, generating response without off-target effects (a drawback of many TAM-depleting agents under clinical study).

Although many studies have explored the depletion of tumor-associated macrophages (TAM) as a therapeutic strategy for solid tumors, currently available compounds suffer from poor efficacy and dose-limiting side effects. Here, we developed a novel TAM-depleting agent (“OximUNO”) that specifically targets CD206+ TAMs and demonstrated efficacy in a triple-negative breast cancer (TNBC) mouse model. OximUNO comprises a star-shaped polyglutamate (St-PGA) decorated with the CD206-targeting peptide mUNO that carries the chemotherapeutic drug doxorubicin (DOX). In the TNBC model, a fluorescently labeled mUNO-decorated St-PGA homed to CD206+ TAMs within primary lesions and metastases. OximUNO exhibited no acute liver or kidney toxicity in vivo. Treatment with OximUNO reduced the progression of primary tumor lesions and pulmonary metastases, significantly diminished the number of CD206+ TAMs and increased the CD8/FOXP3 expression ratio (indicating immunomodulation). Our findings suggest the potential benefit of OximUNO as a TAM-depleting agent for TNBC treatment. Importantly, our studies also represent a novel design of a peptide-targeted St-PGA as a targeted therapeutic nanoconjugate.Significance:A peptide-targeted nanoformulation of DOX exclusively eliminates mannose receptor+ TAMs in breast cancer models, generating response without off-target effects (a drawback of many TAM-depleting agents under clinical study).

One .docx word file containing the following information:Scheme S1. Synthetic approach for St-PGA-OG-mUNO.Scheme S2. Synthetic approach for OximUNO.Video S1. Fifty ns MD trajectory of St-PGA in solution.Table S1. St-PGA-OG-mUNO and St-PGA-OG dye loading, mUNO loading, size and charge.Table S2. OximUNO and St-PGA-DOX drug loading, mUNO loading, size and charge.Table S3. Hepatic and renal toxicity analysed following one dose of OximUNO.Fig. S1. Representative characterisation of St-PGA-OG and St-PGA-OG-mUNO.Fig. S2. The angle used to characterise the rotation of mUNO around PGA.Fig. S3. St-PGA-OG-mUNO shows low hepatic accumulation in the orthotopic TNBC.Fig. S4. Immunostaining showing CD206 expression in the liver compared to the saturated signal of CD206 in the orthotopic 4T1 tumour.Fig. S5. Immunostaining of endogenous IgG indicates leaky tumour vasculature in the orthotopic 4T1 tumours.Fig. S6. Distribution of St-PGA-OG-mUNO in the lung, spleen, sentinel lymph node (SLN) and kidney.Fig. S7. St-PGA-OG-mUNO shows low hepatic accumulation in the experimental metastasis of TNBC.Fig. S8. St-PGA-OG-mUNO shows higher receptor colocalisation than FAM-mUNO.Fig. S9. St-PGA-OG-mUNO shows high homing to M2 TAMs in the orthotopic TNBC but higher hepatic accumulation with a higher dose.Fig. S10. Representative characterisation of St-PGA-DOX and OximUNO.Fig. S11. LC-MS method development for the determination of DOX in drug release studies and stability studies of OximUNO in i.p. fluid and dPBS.Fig. S12. OximUNO shows no signs of hepatic or renal toxicity.Fig. S13. H&E on healthy Balb/c mouse lung.Fig. S14. Localisation of St-PGA-OG/St-PGA-OG-mUNO and tumour vessels.Fig. S15. H&E staining of hearts from the treatment of orthotopic TNBC.Fig. S16. A repetition of OximUNO treatment on the orthotopic TNBC monitoring tumour growth and survival.Fig. S17. Flow cytometry plots for M2 and M1 macrophages.Fig. S18. Flow cytometry gating for cytotoxic T lymphocytes (CTLs).Fig. S19. Flow cytometry gating for T regulatory cells (Tregs). Fig. S20. Flow cytometry analysis showing total cells.Fig. S21. Treatment with OximUNO does not affect splenic macrophages.

One .docx word file containing the following information:Scheme S1. Synthetic approach for St-PGA-OG-mUNO.Scheme S2. Synthetic approach for OximUNO.Video S1. Fifty ns MD trajectory of St-PGA in solution.Table S1. St-PGA-OG-mUNO and St-PGA-OG dye loading, mUNO loading, size and charge.Table S2. OximUNO and St-PGA-DOX drug loading, mUNO loading, size and charge.Table S3. Hepatic and renal toxicity analysed following one dose of OximUNO.Fig. S1. Representative characterisation of St-PGA-OG and St-PGA-OG-mUNO.Fig. S2. The angle used to characterise the rotation of mUNO around PGA.Fig. S3. St-PGA-OG-mUNO shows low hepatic accumulation in the orthotopic TNBC.Fig. S4. Immunostaining showing CD206 expression in the liver compared to the saturated signal of CD206 in the orthotopic 4T1 tumour.Fig. S5. Immunostaining of endogenous IgG indicates leaky tumour vasculature in the orthotopic 4T1 tumours.Fig. S6. Distribution of St-PGA-OG-mUNO in the lung, spleen, sentinel lymph node (SLN) and kidney.Fig. S7. St-PGA-OG-mUNO shows low hepatic accumulation in the experimental metastasis of TNBC.Fig. S8. St-PGA-OG-mUNO shows higher receptor colocalisation than FAM-mUNO.Fig. S9. St-PGA-OG-mUNO shows high homing to M2 TAMs in the orthotopic TNBC but higher hepatic accumulation with a higher dose.Fig. S10. Representative characterisation of St-PGA-DOX and OximUNO.Fig. S11. LC-MS method development for the determination of DOX in drug release studies and stability studies of OximUNO in i.p. fluid and dPBS.Fig. S12. OximUNO shows no signs of hepatic or renal toxicity.Fig. S13. H&E on healthy Balb/c mouse lung.Fig. S14. Localisation of St-PGA-OG/St-PGA-OG-mUNO and tumour vessels.Fig. S15. H&E staining of hearts from the treatment of orthotopic TNBC.Fig. S16. A repetition of OximUNO treatment on the orthotopic TNBC monitoring tumour growth and survival.Fig. S17. Flow cytometry plots for M2 and M1 macrophages.Fig. S18. Flow cytometry gating for cytotoxic T lymphocytes (CTLs).Fig. S19. Flow cytometry gating for T regulatory cells (Tregs). Fig. S20. Flow cytometry analysis showing total cells.Fig. S21. Treatment with OximUNO does not affect splenic macrophages.

Although many studies have explored the depletion of tumor-associated macrophages (TAM) as a therapeutic strategy for solid tumors, currently available compounds suffer from poor efficacy and dose-limiting side effects. Here, we developed a novel TAM-depleting agent (“OximUNO”) that specifically targets CD206+ TAMs and demonstrated efficacy in a triple-negative breast cancer (TNBC) mouse model. OximUNO comprises a star-shaped polyglutamate (St-PGA) decorated with the CD206-targeting peptide mUNO that carries the chemotherapeutic drug doxorubicin (DOX). In the TNBC model, a fluorescently labeled mUNO-decorated St-PGA homed to CD206+ TAMs within primary lesions and metastases. OximUNO exhibited no acute liver or kidney toxicity in vivo. Treatment with OximUNO reduced the progression of primary tumor lesions and pulmonary metastases, significantly diminished the number of CD206+ TAMs and increased the CD8/FOXP3 expression ratio (indicating immunomodulation). Our findings suggest the potential benefit of OximUNO as a TAM-depleting agent for TNBC treatment. Importantly, our studies also represent a novel design of a peptide-targeted St-PGA as a targeted therapeutic nanoconjugate.Significance: A peptide-targeted nanoformulation of DOX exclusively eliminates mannose receptor+ TAMs in breast cancer models, generating response without off-target effects (a drawback of many TAM-depleting agents under clinical study).

Although many studies have explored the depletion of tumour-associated macrophages (TAMs) as a therapeutic strategy for solid tumours, currently available compounds suffer from poor efficacy and dose-limiting side effects. Here, we developed a novel TAM-depleting agent ("OximUNO") that specifically targets CD206⁺ TAMs and demonstrated efficacy in triple negative breast cancer (TNBC) mouse models. OximUNO comprises a star-shaped polyglutamate (St-PGA) decorated with the CD206-targeting peptide mUNO that carries the chemotherapeutic drug doxorubicin (DOX). In TNBC models, a fluorescently-labelled mUNO-decorated St-PGA homed to CD206⁺ TAMs within primary lesions and metastases. OximUNO exhibited no acute liver or kidney toxicity in vivo. Treatment with OximUNO reduced the progression of primary tumour lesions and pulmonary metastases, significantly diminished the number of CD206⁺ TAMs and increased the CD8/FOXP3 expression ratio (demonstrating immunostimulation). Our findings suggest the potential benefit of OximUNO as a TAM-depleting agent for TNBC treatment. Importantly, our studies also represent the first report of a peptide-targeted St-PGA as a targeted therapeutic nanoconjugate.