Figure S5
A, B. Flow cytometric analysis of MOC1esc1_Ctrl and MOC1esc1_CCL5 tumors harvested on day 16 post tumor inoculation. Number of cells per tumor mg
are shown. (n=6 for esc1_Ctrl, n=8 for esc1_CCL5, representative data of two independent experiments.)
C. Flow cytometric analysis of MOC1esc1_Ctrl and MOC1esc1_CCL5 DLN harvested on day 16 post tumor inoculation. (Left panel, n=12 for Ctrl, n=14 for
esc1_CCL5, pooled data from two independent experiments. Right panel, n=18 for Ctrl, n=20 for esc1_CCL5. Pooled data from three independent experiments.)
D, E. Flow cytometric analysis of MOC1esc1_Ctrl and MOC1esc1_CCL5 DLN harvested on day 16 post tumor inoculation. (n= 6 for Ctrl, n=8 for CCL5,
representative data of two independent experiments.)
F. Total cells (5*105cells/well) isolated from MOC1esc1_Ctrl or MOC1esc1_CCL5 DLNs were stimulated with indicated peptides for 48 hours and evaluated
by IFN-γ ELISA. PC; (positive control (PMA + ionomycin)), NC; negative control (no peptides). (Left panel, n=8. Right panel, n=3, representative data of
two independent experiments.)
G. Flow cytometric analysis of MOC2_Ctrl and MOC2_CCL5 tumors harvested on day 10 post tumor inoculation. (n=8.)
ARTICLE ABSTRACT
Neoadjuvant anti-PD1 (aPD1) therapies are being explored in surgically resectable head and neck squamous cell carcinoma (HNSCC). Encouraging responses have been observed, but further insights into the mechanisms underlying resistance and approaches to improve responses are needed.
We integrated data from syngeneic mouse oral carcinoma (MOC) models and neoadjuvant pembrolizumab HNSCC patient tumor RNA-sequencing data to explore the mechanism of aPD1 resistance. Tumors and tumor-draining lymph nodes (DLN) from MOC models were analyzed for antigen-specific priming. CCL5 expression was enforced in an aPD1-resistant model.
An aPD1-resistant mouse model showed poor priming in the tumor DLN due to type 1 conventional dendritic cell (cDC1) dysfunction, which correlated with exhausted and poorly responsive antigen-specific T cells. Tumor microenvironment analysis also showed decreased cDC1 in aPD1-resistant tumors compared with sensitive tumors. Following neoadjuvant aPD1 therapy, pathologic responses in patients also positively correlated with baseline transcriptomic cDC1 signatures. In an aPD1-resistant model, intratumoral cDC1 vaccine was sufficient to restore aPD1 response by enhancing T-cell infiltration and increasing antigen-specific responses with improved tumor control. Mechanistically, CCL5 expression significantly correlated with neoadjuvant aPD1 response and enforced expression of CCL5 in an aPD1-resistant model, enhanced cDC1 tumor infiltration, restored antigen-specific responses, and recovered sensitivity to aPD1 treatment.
These data highlight the contribution of tumor-infiltrating cDC1 in HNSCC aPD1 response and approaches to enhance cDC1 infiltration and function that may circumvent aPD1 resistance in patients with HNSCC.
