HPV8-induced STAT3 activation led keratinocyte stem cell expansion in human actinic keratoses
Huw J. Morgan, Carlotta Olivero, Boris Y. Shorning, Alex Gibbs, Alexandra L. Phillips, Lokapriya Ananthan, Annabelle Xiao Hui Lim, Licia Martuscelli, Cinzia Borgogna, Marco De Andrea, Martin Hufbauer, Richard Goodwin, Baki Akgül, Marisa Gariglio, Girish K. Patel
Huw J. Morgan, Carlotta Olivero, Boris Y. Shorning, Alex Gibbs, Alexandra L. Phillips, Lokapriya Ananthan, Annabelle Xiao Hui Lim, Licia Martuscelli, Cinzia Borgogna, Marco De Andrea, Martin Hufbauer, Richard Goodwin, Baki Akgül, Marisa Gariglio, Girish K. Patel
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Research Article Cell biology Stem cells

HPV8-induced STAT3 activation led keratinocyte stem cell expansion in human actinic keratoses

Abstract

Despite epidermal turnover, the skin is host to a complex array of microbes, including viruses, such as HPV, which must infect and manipulate skin keratinocyte stem cells (KSCs) to survive. This crosstalk between the virome and KSC populations remains largely unknown. Here, we investigated the effect of HPV8 on KSCs using various mouse models. We observed that the HPV8 early region gene E6 specifically caused Lrig1+ hair follicle junctional zone KSC proliferation and expansion, which would facilitate viral transmission. Within Lrig1+ KSCs specifically, HPV8 E6 bound intracellular p300 to phosphorylate the STAT3 transcriptional regulatory node. This induced ΔNp63 expression, resulting in KSC expansion into the overlying epidermis. HPV8 was associated with 70% of human actinic keratoses. Together, these results define the “hit-and-run” mechanism for HPV8 in human actinic keratosis as an expansion of KSCs, which lack melanosome protection and are thus susceptible to sun light–induced malignant transformation.

Authors

Huw J. Morgan, Carlotta Olivero, Boris Y. Shorning, Alex Gibbs, Alexandra L. Phillips, Lokapriya Ananthan, Annabelle Xiao Hui Lim, Licia Martuscelli, Cinzia Borgogna, Marco De Andrea, Martin Hufbauer, Richard Goodwin, Baki Akgül, Marisa Gariglio, Girish K. Patel

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Figure 4

Lrig1+ hair follicle junctional zone KSC progeny retain KSCs.

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Lrig1+ hair follicle junctional zone KSC progeny retain KSCs. (A) Immuno...
(A) Immunoblot of pSTAT3 Y705 and S727, with TATA-Box binding protein (TBP) control (n = 3). (B) CLSM of dorsal back skin for lineage tracing of Lrig1CreERT2:R26RConfetti:WT and Lrig1CreERT2:R26RConfetti:HPV8-E6tg progeny, 4 weeks after Cre activation. Scale bar: 40 μm. (C) Enumerated Lrig1+Confetti+ cells and their progeny Lrig1–Confetti+ flow-sorted cell populations from Lrig1CreERT2:R26RConfetti:HPV8-E6tg and Lrig1CreERT2:R26RConfetti:WT mice (n = 25 total). (D) Immunoblot of Lrig1+Confetti+ cells and their progeny Lrig1–Confetti+ flow-sorted cell populations (n = 3). (E) Venn diagram showing shared DEGs from Confetti HPV8 E6 versus Confetti WT comparisons for Lrig1+Confetti+ and Lrig1– Confetti+ populations (see Supplemental Table 2). (F) GSEA for STAT3- and EMT-associated gene signatures in DEGs from Lrig1+Confetti+ transcriptomic comparison of Confetti HPV8 E6 versus Confetti WT analysis. (G) GSEA for STAT3-, EMT- and differentiation-associated gene signatures in DEGs from Lrig1–Confetti+ transcriptomic comparison of Confetti HPV8 E6 versus Confetti WT analysis. (H) GSEA for STAT3- and EMT-associated gene signatures in DEGs from Confetti HPV8 E6 transcriptomic comparison of Lrig1+Confetti+ and Lrig1–Confetti+ population analysis. See also Supplemental Figure 3. (I) qPCR of RNA from flow-sorted cell isolates as in C for STAT3-regulated genes (n = 3). (J) CFE of 2,500 flow-sorted Lrig1+Confetti+ and Lrig1–Confetti+ flow-sorted cell populations from Lrig1CreERT2:R26RConfetti:HPV8-E6tg (n = 4). See also Supplemental Figure 3. Statistical tests included (A, D, and I) 1-way ANOVA and (C and J) 2-tailed Student’s t test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.