044 Metatranscriptomics reveals association of α-, β-, and γ-HPVs with typical epidermodysplasia verruciformis in a large cohort of patients with CIB1, TMC6, or TMC8 mutations

      Cutaneous human papillomavirus (HPV) infection typically manifests with isolated warts. However, some patients in familial clustering develop extensive and protracted HPV infections, primarily the β-HPV types 5 and 8, with distinct cutaneous findings. This clinical entity, epidermodysplasia verruciformis (EV), with autosomal recessive inheritance, is characterized by numerous cutaneous flat warts in childhood, which progress into squamous cell carcinomas later in life. The “typical” form of EV, not vulnerable to other infections, is caused by mutations in CIB1, TMC6, or TMC8, which impair keratinocyte-intrinsic immunity to β-HPV infection. Mutations in other genes related to T-cell development or function, have been associated with “atypical” EV in patients with other infections. We developed a whole-transcriptome sequencing-based method on RNA isolated from skin biopsies for concomitant detection of viral and human genetic determinants of cutaneous wart lesions in a cohort of 50 EV patients. This method, VirPy, can detect 926 viruses, including more than 400 HPVs, and the corresponding human mutations. Nine distinct mutations in TMC8 (n=2), TMC6 (n= 5) and CIB1 (n=2) in 12 distinct families, including 14 patients were detected. The most predominant HPV in this cohort was HPV14. In addition, the RNA-seq data were examined for variant detection and prioritization, pathogenicity confirmation, and RNA expression profiling. Besides, we identified a total of 20 different HPVs including 16 β-, three α- and one γ-HPV(HPV128) in a patient with TMC8 mutation. In summary, the utilization of RNA-Seq as a first-tier diagnostic method allowed us to simultaneously profile the transcriptome of host for mutation detection and exploring the consequences of variants of unknown significance as well as to profile the cutaneous virome of EV patients.