CD47 blockade enhances immunoradiotherapy response in head and neck squamous cell carcinoma

Monther, A.; Al-Msari, R.; Saddawi-Konefka, R.; Fassardi, S.; Tang, C.; Philips, C.; Sen, P.; Mohammadzadeh, P.; Decker, K.; Miyauchi, S.; Roy, S.; Jones, R.; Gutkind, S.; Sharabi, A.; Califano, J.

doi:10.1101/2025.06.29.662015

bioRxiv · 10.1101/2025.06.29.662015

CD47 blockade enhances immunoradiotherapy response in head and neck squamous cell carcinoma

Monther, A.·Al-Msari, R.·Saddawi-Konefka, R.·Fassardi, S.·Tang, C.·Philips, C.·Sen, P.·Mohammadzadeh, P.·Decker, K.·Miyauchi, S.·Roy, S.·Jones, R.·Gutkind, S.·Sharabi, A.·Califano, J.

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Abstract

Head and neck squamous cell carcinoma (HNSCC) is a significant cause of morbidity and mortality worldwide, with limited treatment options for patients with locally advanced disease. CD47 immune checkpoint inhibitors have been used to block the CD47/SIRPa interaction that inhibits antigen-presenting cell phagocytosis, thereby enhancing antigen presentation to cytotoxic T-cells, and have shown promise in combination with anti-PD1 immunotherapy in tumors, including recurrent/metastatic HNSCC. We found that CD47 expression is associated with poor prognosis in HNSCC and explored the anti-tumor activity of CD47 blockade in combination with anti-PD1 and lymphatic-sparing radiotherapy in a locally advanced HNSCC model. Using the 4MOSC1 orthotopic, syngeneic murine model of HPV-negative HNSCC, treatment with an engineered CD47-blocking SIRP fusion protein (ALX301) similarly induced complete tumor regression in combination with anti-PD1, and a partial response as a standalone therapeutic. An anti-PD1 immune checkpoint inhibitor in a CD47-null tumor background led to complete tumor regression confirming a key role for CD47 in tumor immunity. Anti-CD47 treated mice demonstrated increased MHC-II expression on dendritic cells within the tumor and upregulation of CD86 co-stimulatory molecule on dendritic cells within the tumor, sentinel lymph nodes, and contralateral lymph nodes. Combination ALX301 and anti-PD1 treatment in an anti-PD1 resistant 4MOSC2 model demonstrated significant tumor regression, enhanced survivability, improved response with neoadjuvant radiotherapy, and greater retention of CD8+ T-cells within the tumor microenvironment. Notably, T-cell receptor sequencing revealed increased shared clonality between the tumor and sentinel lymph nodes of anti-CD47 treated mice. These data demonstrate that a combination of CD47 blockade and anti-PD1 therapy enhances tumor antigen presentation and immune cell infiltration, while further improving anti-tumor responses in combination with tumor-targeted radiotherapy. This study provides support for the rational design of combinatorial immunoradiotherapy, using anti-CD47 inhibitors and anti-PD1 therapy, in a clinical trial targeting locally advanced HPV-negative HNSCC.

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Monther, A., Al-Msari, R., Saddawi-Konefka, R., Fassardi, S., Tang, C., Philips, C., Sen, P., Mohammadzadeh, P., Decker, K., Miyauchi, S., Roy, S., Jones, R., Gutkind, S., Sharabi, A., Califano, J.. 2025-06-30. CD47 blockade enhances immunoradiotherapy response in head and neck squamous cell carcinoma. https://doi.org/10.1101/2025.06.29.662015

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