Background: Patients with non-small cell lung cancer (NSCLC) harboring PTEN mutations have a worse prognosis of immunotherapy in research with small sample size and case reports. However, the role of PTEN and TP53 co-mutations in immunotherapy of NSCLC has not been fully studied. Methods: Patients with NSCLC from our dataset were enrolled in this study. For the genomic profiling, only the pathogenic mutations and likely pathogenic mutations in clinical significance were rolled. The efficiency of immunotherapy was analyzed in a public cohort of 240 NSCLC patients by Rizvi. Patients with EGFR/ALK mutation were excluded in our analysis. Results: In our dataset, 1,428 patients were included in the analysis. 25 (1.75%) patients have PTEN mutations and 429 (30.04%) patients have TP53 mutations. In all patients with PTEN mutations, 84% with TP53 co-mutations. Median tumor mutational burden (TMB) was 12.29 mutations/MB (range, 3.35-36.87) and 6.98mutations/MB (range, 0.56-9.50) in patients harboring PTEN mutations with TP53 co-mutations and without TP53 co-mutations, respectively (p= 0.02). In 240 NSCLC patients of Rizvi, the overall response rate (ORR) of the PTEN and TP53 co-mutations group, the PTEN mutations without TP53 mutations group and PTEN wild-type group was 33.33%, 0% and 20.26%, respectively. The disease control rate (DCR) was 83.33%, 0% and 54.74%, respectively. The median progression-free survival (PFS) in the PTEN and TP53 co-mutations group was the longest (7.5 months), while the median PFS in the PTEN mutations without TP53 mutations group was only 2.5 months. However, No significant differences of PFS were observed. Conclusions: Our findings suggested that patients with NSCLC harboring PTEN and TP53 co-mutations might benefit more from immunotherapy compare to those harboring PTEN mutation without TP53 mutations and PTEN wild-type. TMB may be an important factor that lead to the results. Further studies need to be explore in cohorts with larger sample sizes.