Head and neck squamous cell carcinoma (HNSCC) is the sixth most common type of cancer worldwide. HNSCC is highly aggressive and arises from the epithelial mucosal layer of the aerodigestive tract. Despite new treatment approaches, the 5-year survival rate remains at 40%, highlighting the need for new therapeutic strategies. To exploit the inherent immunosurveillance and killing activity of natural killer (NK) cells paired with their potential for an allogeneic off-the-shelf product, we used chimeric antigen receptors (CAR) to redirect NK cell cytotoxicity by targeting tumor-associated antigens HER1 and CD44v6. During an initial screening of third-generation anti-HER1 CARs, we selected two candidates based on stable CAR expression and lack of tonic signaling. Anti-HER1 CAR-NK-92 cells effectively killed HNSCC cell lines (SCC-4 and SCC-25) and primary HNSCC (pHNSCC) cells in a flow cytometry based cytotoxicity assay. Characterization of pHNSCC samples prior to coculture with CAR-NK cells revealed high percentages of CD45⁺ immune cells, including T cells, B cells, NK cells and macrophages. To prevent contamination with fibroblast and immune cells, an ex vivo expansion protocol that includes a sorting step for epithelial marker EpCAM⁺ cells was established. Examination of the remaining primary HNSCC cells after coculture with our CAR-NK cells showed loss of HER1⁺ and HER1⁺CD44v6⁺ populations with an expansion of CD44v6⁺ populations. In summary, our data indicate the need for dual-targeted CAR-NK based therapies that can also be combined with currently available treatment regimens, including radiotherapy and checkpoint inhibition to improve treatment outcomes and survival rates for HNSCC.