Tobacco smoking-associated alterations in the immune microenvironment of squamous cell carcinomas

Background: Tobacco smoking creates DNA damage, inducing mutations and potentially altering the tumor immune microenvironment. These types of genetic and immune microenvironment alterations are critical factors known to affect tumor response to immunotherapy. Here we analyze the association between the mutational signature of tobacco smoking, tumor mutational load, and metrics of immune activity in squamous cell carcinomas arising in the head and neck and lung. Methods: Using RNA and DNA sequencing data from The Cancer Genome Atlas head and neck (HNSC; n ¼ 287) and lung (LUSC; n ¼ 130) squamous cell carcinoma data sets and two independent gene expression data sets (HNSC, n ¼ 136; LUSC, n ¼ 75), we examined associations between the mutational smoking signature, mutation count, immune cell infiltration, cytolytic activity, and interferon-c signaling. Results: An increasing mutational smoking signature was associated with statistically significantly increased overall mutational load in both HNSC (q ¼ .33, P ¼ 1.01 10⁷) and LUSC (q ¼ .49, P ¼ 2.80 10⁹). In HNSC, a higher mutational smoking signature was associated with lower levels of immune infiltration (q ¼ -.37, P ¼ 1.29 10¹⁰), cytolytic activity (q ¼ -.28, P ¼ 4.07 10⁶), and interferon-c pathway signaling (q ¼ .39, P ¼ 3.20 10¹¹). In LUSC, these associations were reversed (q ¼ .19, P ¼ .03; q ¼ .20, P ¼ .02; and q ¼ .18, P ¼ .047, respectively). Differentially expressed genes between smoking-high and smoking-low tumors revealed broad tobacco-induced immunosuppression in HNSC, in contrast to a tumor-inflamed microenvironment in smokers with LUSC. Conclusions: In squamous cell carcinomas, the genetic smoking signature is associated with higher mutational load, but variable effects on tumor immunity can occur, depending on anatomic site. In HNSC, smoking is predominantly immunosuppressive; in LUSC, more pro-inflammatory. Both tumor mutation load and immune microenvironment affect clinical response to immunotherapy. Thus, the mutational smoking signature is likely to have relevance for immunothera-peutic investigation in smoking-associated cancers.