The HPV and p63 Status in Penile Cancer Are Linked with the Infiltration and Therapeutic Availability of Neutrophils

Abstract

Squamous penile cancer displays a rare human papillomavirus (HPV)–associated tumor entity. Investigations on the molecular pathogenesis of HPV-driven penile cancer are impaired by the rareness of clinical specimens and, in particular, are missing relevant cell culture models. Here, we identified in HPV-positive penile cancer cell lines that HPV16 oncoproteins control TP63 expression by modulating critical regulators, while integration into the TP63 open reading frame facilitates oncogene expression. The resulting feed-forward loop leads to elevated p63 levels that in turn enhance the release of the neutrophil-recruiting chemokine CXCL8. Remarkably, elevated CXCL8 amounts lead to the increased surface exposition of the Fc receptor of human IgA antibodies, FcαRI, on neutrophils and correlated with a higher susceptibility to antibody-dependent neutrophil-mediated cytotoxicity (ADCC) using an EGFR-specific IgA2 antibody. IHC staining of tissue microarrays proved that elevated expression of p63 together with neutrophil infiltration were significantly more frequent in HPV-positive penile cancer displaying a higher tumor grade. In summary, we identified a promising marker profile of patients with penile cancer at higher risk for worse prognosis. However, these patients may benefit from immunotherapeutic approaches efficiently engaging neutrophils for tumor cell killing.

The prominent role of the S100A8/S100A9-CD147 axis in the progression of penile cancer

“…Current epidemiologic data on PSCC imply a rising incidence of this HPV-associated malignant disease ( 33 – 35 ). These data further underline the need for promising markers, especially of MDSCs, because our recently published data already suggested the involvement of myeloid cells during HPV-driven penile carcinogenesis ( 16 , 36 ). Tissue microarrays (TMAs) with a high number of clinical PeCa specimens with matching clinic-pathological data were used to identify a promising candidate marker profile of patients at a higher risk for lymph node involvement.…”

“…The TMA cohort consists of patients derived from Russia and Germany between 1992 and 2015. Data on clinical outcome and HPV status were published previously ( 15 , 16 ). Briefly, DNA was isolated from formalin-fixed paraffin-embedded (FFPE) tissue sections by the QIAamp DNA FFPE Tissue Kit (Qiagen, Hilden, Germany) following the manufacturer’s protocol, and the HPV PCR was conducted using the GP5+/6+ primers as described previously ( 15 , 16 ).…”

“…Data on clinical outcome and HPV status were published previously ( 15 , 16 ). Briefly, DNA was isolated from formalin-fixed paraffin-embedded (FFPE) tissue sections by the QIAamp DNA FFPE Tissue Kit (Qiagen, Hilden, Germany) following the manufacturer’s protocol, and the HPV PCR was conducted using the GP5+/6+ primers as described previously ( 15 , 16 ). HPV status was further determined by p16 INK4A immunohistochemistry (IHC), a surrogate marker for active HPV oncoprotein–driven transformation, using a published protocol ( 15 , 16 ).…”

“…Briefly, DNA was isolated from formalin-fixed paraffin-embedded (FFPE) tissue sections by the QIAamp DNA FFPE Tissue Kit (Qiagen, Hilden, Germany) following the manufacturer’s protocol, and the HPV PCR was conducted using the GP5+/6+ primers as described previously ( 15 , 16 ). HPV status was further determined by p16 INK4A immunohistochemistry (IHC), a surrogate marker for active HPV oncoprotein–driven transformation, using a published protocol ( 15 , 16 ). HPV status was considered as positive in case of positive PCR and IHC with subsequent genotyping using INNO-LiPA genotyping extra II (Fujirebio Germany GmbH, Hannover, Germany), revealing HPV16, HPV18, and HPV59 in 27 (93.1%), one (3.45%), and one (3.45%) of the 29 cases, respectively.…”

“…We recently described CD15, a surface protein of neutrophils, as a promising candidate and identified an HPV active p63 high CXCL8 high axis as a possible underlying mechanism that drives neutrophil infiltration ( 16 ). Subsequent inclusion of clinic-pathological data revealed that especially HPV + p63 + CD15 + PeCa was characterized by dedifferentiation, invasion, and metastases.…”