Development Of Non-melanoma Skin Cancer Research Articles

CYLD Inhibits the Development of Skin Squamous Cell Tumors in Immunocompetent Mice.

Cylindromatosis (CYLD) is a deubiquitinase (DUB) enzyme that was initially characterized as a tumor suppressor of adnexal skin tumors in patients with CYLD syndrome. Later, it was also shown that the expression of functionally inactive mutated forms of CYLD promoted tumor development and progression of non-melanoma skin cancer (NMSC). However, the ability of wild-type CYLD to inhibit skin tumorigenesis in vivo in immunocompetent mice has not been proved. Herein, we generated transgenic mice that express the wild type form of CYLD under the control of the keratin 5 (K5) promoter (K5-CYLDwt mice) and analyzed the skin properties of these transgenic mice by WB and immunohistochemistry, studied the survival and proliferating characteristics of primary keratinocytes, and performed chemical skin carcinogenesis experiments. As a result, we found a reduced activation of the nuclear factor kappa B (NF-κB) pathway in the skin of K5-CYLDwt mice in response to tumor necrosis factor-α (TNF-α); accordingly, when subjected to insults, K5-CYLDwt keratinocytes are prone to apoptosis and are protected from excessive hyperproliferation. Skin carcinogenesis assays showed inhibition of tumor development in K5-CYLDwt mice. As a mechanism of this tumor suppressor activity, we found that a moderate increase in CYLD expression levels reduced NF-κB activation, which favored the differentiation of tumor epidermal cells and inhibited its proliferation; moreover, it decreased tumor angiogenesis and inflammation. Altogether, our results suggest that increased levels of CYLD may be useful for anti-skin cancer therapy.

249 Increased susceptibility to sunburn occurs during the first three years of hydrochlorothiazide use: Results from the National Health and Nutrition Examination Survey, 2009-2018

Hydrochlorothiazide (HCTZ) has been associated with increased photosensitivity and development of non-melanoma skin cancer. Given the Food and Drug Administration’s recommendation of increased sun protection in patients taking HCTZ, we investigated the relationship between HCTZ use, photoprotection behaviors, and sunburn using the National Health and Nutrition Examination Survey (NHANES). We combined NHANES data from 2009-2018 (931 taking HCTZ, 18148 not taking HCTZ) and used multiple logistic regression to calculate adjusted odds ratios (aOR) of staying in shade, wearing long sleeves, and using sunscreen. We used quasi-Poisson regression with linear spline to model incidence rate ratios (IRR) of sunburns in the past year associated with duration of HCTZ use. Models were adjusted for survey cycle, sex, age, race/ethnicity, education level, income to poverty ratio, history of skin cancer, and time spent outdoors. We controlled for photoprotection behaviors when examining sunburn as the outcome. Current HCTZ use was not associated with change in odds of frequently staying in shade (aOR 1.11, CI 0.93 – 1.32, P=0.253), wearing long sleeves (aOR 1.38, CI 0.99 – 1.91, P=0.063), or using sunscreen (aOR 1.11, CI 0.89 – 1.39, P=0.363). Among participants who took HCTZ for 3 years or less, each additional year of HCTZ use was associated with a 38% increase in incidence rate of sunburns in the past year (IRR 1.38, CI 1.12 – 1.70, p=0.004). Among participants who took HCTZ for more than 3 years, no statistically significant association between duration of HCTZ use and incidence rate of sunburns was found (IRR 0.98, CI 0.93 – 1.04, p=0.526). In summary, we found areas for improvement in sun protection among patients taking HCTZ and increased susceptibility to sunburn during the first 3 years of HCTZ use. Education on photoprotection methods, especially with initiation of HCTZ, is important in this population to prevent photodamage and the development of skin cancer.

073 A non-invasive genomic test for early assessment of UV damage in human skin

UV-exposure leads to mutations in skin cells, which may clonally expand and ultimately turn into cancerous lesions ('field cancerization'). Early detection of low-frequency cancer-causing UV-signature mutations in still normal-appearing skin remains a challenge. We have developed a non-invasive genomic test capable of quantifiably detecting UV-signature mutations with high sensitivity to change this paradigm. This genomic test combines DermTech's non-invasive skin sample collection technology 'adhesive skin collection kits' with an ultrasensitive and multiplexed MALDI-TOF mass spectrometry-based mutation detection platform, 'the MassARRAY' from Agena Bioscience. The test includes over two dozen DNA driver mutations in genes such as TP53, CDKN2A, NOTCH1, and NOTCH2, known to be associated with the development of non-melanoma skin cancers. In a proof-of-concept study, contralaterally collected skin samples from sun-exposed body sites in subjects of varying race, age, and gender were tested. UV-signature mutations were detected in sun-exposed skin, with the numbers of mutations increasing with the subjects' age. In our reference control neonatal keratinocytes, no UV signature mutations were detected. This genomic test demonstrates reliable detection of UV-signature mutations in sun-exposed skin samples collected using DermTech's non-invasive skin sample collection technology and offers the opportunity to non-invasively assess UV-damage of skin that lacks visual elements of UV damage. Additional studies will help understand the correlation of UV-damage detected in normal-appearing skin to future non-melanoma skin cancer risk. We envision this proactive genomic approach to measure UV damage to change how photoaging is assessed and treated.

Adhesion Molecules in Non-melanoma Skin Cancers: A Comprehensive Review.

Basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) are the most frequently diagnosed cancers, generating significant medical and financial problems. Cutaneous carcinogenesis is a very complex process characterized by genetic and molecular alterations, and mediated by various proteins and pathways. Cell adhesion molecules (CAMs) are transmembrane proteins responsible for cell-to-cell and cell-to-extracellular matrix adhesion, engaged in all steps of tumor progression. Based on their structures they are divided into five major groups: cadherins, integrins, selectins, immunoglobulins and CD44 family. Cadherins, integrins and CD44 are the most studied in the context of non-melanoma skin cancers. The differences in expression of adhesion molecules may be related to the invasiveness of these tumors, through the loss of tissue integrity, neovascularization and alterations in intercellular signaling processes. In this article, each group of CAMs is briefly described and the present knowledge on their role in the development of non-melanoma skin cancers is summarized.

Vitamin D Receptor Polymorphisms and Non-Melanoma Skin Cancer Risk: A Case-Control Study.

Exposure to sunlight is the major source of vitamin D and the main environmental cause of non-melanocytic skin cancers. Vitamin D, partly mediated through the vitamin D receptor (VDR), has potential therapeutic applications in skin cancer. The aim of this study was to investigate the association of BsmI and ApaI VDR polymorphisms among patients with non-melanoma cancers and controls. An observational case-control study was conducted in a sample of 154 subjects. We observed no significant effects between these polymorphisms and skin cancer risk. When stratified for gender, GG and AG BsmI polymorphisms significantly increased the risk of basal cell carcinomas in males. In relation to ApaI, all three polymorphisms significantly increased the risk of basal cell carcinoma in males. When stratified for age, we found that being 70 years of age or younger was a protective factor against both skin cancers. Being a female and 70 years old or younger was a protective factor for basal cell carcinoma. A comparison of the frequencies of the VDR genotypes in patients older than 70 years vs. 70 years or younger also revealed age-dependent variations in patients with non-melanoma skin cancer. Our study suggests a role for VDR polymorphisms in non-melanoma skin cancer development.

Do Diet and Lifestyles Play a Role in the Pathogenesis of NMSCs?

Background and Aims: Literature highlights the role of risk factors like age, body mass index (BMI), tobacco smoking, alcohol intake and diet in the pathogenesis of several cancer types but little is known for non-melanoma skin cancers (NMSC). The aim of this epidemiological study was to evaluate the correlation between modifiable risk factors (BMI, metabolic panel, diet, lifestyle, medical history) and not modifiable risk factors (gender, age) and NMSC development. Methods: From February 2018 to September 2019, 162 patients affected by NMSC were compared to a group of 167 controls. A univariate and multivariate analysis was conducted to elaborate the data collected through face-to-face interviews. Results: While our evidence did not always reach statistical significance, NMSC study group patients exhibited high rates of analyzed risk factors (male gender aging over 55 years, high BMI, reduced physical activity) compared to the control group. Conclusions: Our study indicates that practicing more than 30 min of physical activity daily could be a protective factor against the NMSC onset. Other risk factors were not correlated with NMSC, but more evidence is needed to establish a possible link.

Gene expression is stable in a complete CIB1 knockout keratinocyte model

Epidermodysplasia verruciformis (EV) is a genodermatosis characterized by the inability of keratinocytes to control cutaneous β-HPV infection and a high risk for non-melanoma skin cancer (NMSC). Bi-allelic loss of function variants in TMC6, TMC8, and CIB1 predispose to EV. The correlation between these proteins and β-HPV infection is unclear. Its elucidation will advance the understanding of HPV control in human keratinocytes and development of NMSC. We generated a cell culture model by CRISPR/Cas9-mediated deletion of CIB1 to study the function of CIB1 in keratinocytes. Nine CIB1 knockout and nine mock control clones were generated originating from a human keratinocyte line. We observed small changes in gene expression as a result of CIB1 knockout, which is consistent with the clearly defined phenotype of EV patients. This suggests that the function of human CIB1 in keratinocytes is limited and involves the restriction of β-HPV. The presented model is useful to investigate CIB1 interaction with β-HPV in future studies.

628 Evidence that wounding of geriatric skin which upregulates IGF-1 levels protects against both abnormal carcinogenic UVB responses as well as from the development of non-melanoma skin cancer

628 Evidence that wounding of geriatric skin which upregulates IGF-1 levels protects against both abnormal carcinogenic UVB responses as well as from the development of non-melanoma skin cancer

155 A biomarker function of HMGA2 in cutaneous squamous cell carcinoma development

The high mobility group AT-hook 2 (HMGA2) gene encodes a transcription factor that is expressed during embryonic development but down-regulated in adult tissues. Re-expression of HMGA2 in adult tissues is often associated with both benign and malignant tumor formation. HMGA2 has been identified as a biomarker of melanoma progression and prognosis, but its role in non-melanoma skin cancer development remains controversial. In this study, we measured HMGA2 expression in normal human keratinocytes and found a significant decrease in HMGA2 mRNA expression in adult keratinocytes compared to neonatal keratinocytes. HMGA2 is highly expressed in human cutaneous squamous cell carcinoma (SCC) cell lines and primary human SCC tumors, but not detected in adjacent normal skin. In mouse skin, Hmga2 has been found to be translocated from the cell membrane into the nucleus during DMBA/TPA-induced skin tumorigenesis. While Hmga2 expression is absent in the hair follicle in non-UV-irradiated mouse, UV irradiation increased Hmga2 expression in both the epidermis and hair follicles. In agreement, we found enhanced Hmga2 expression in UV-induced mouse skin SCCs and also Hmga2 reactivation in the dermis from UV-irradiated mice. Furthermore, oncogene depletion such as FOXM1 and TRIP13 in human SCC cell lines using CRISPR/Cas9 decreased the expression of HMGA2 coupled with decreased cell proliferation and survival. Taken together, these results demonstrate that the upregulation of HMGA2 is an important biomarker of skin SCC tumorigenesis. Moreover, understanding of the mechanism for HMGA2 overexpression in skin hair follicle cells can develop the new drug for skin cancer treatment.

The Inflammasome Adaptor ASC Delays UV-Induced Skin Tumorigenesis in Beta HPV38 E6 and E7 Transgenic Mice

Non-melanoma skin cancer is the most common cancer in fair-skinned adults and comprises basal cell carcinoma and squamous cell carcinoma (SCC). Its incidence reaches 1 of 100 person-year in Australia (Lomas et al., 2012) and is still on the rise worldwide (Madan et al., 2010). The development of non-melanoma skin cancer depends on genetic and environmental factors. Particularly, there is clear evidence that cumulative sun exposure contributes to the development of non-melanoma skin cancer (Ananthaswamy et al., 1997; Madan et al., 2010).

Abstract IA15: Role of human papillomaviruses in carcinogenesis

Abstract Infectious agents represent a major group of risk factors for cancer development and contribute to about 15% of human cancers worldwide. Six viruses and one bacterium, i.e., human papillomavirus (HPV), hepatitis C virus (HCV), hepatitis B virus, human T-lymphotropic virus type I (HTLV-1), Epstein-Barr virus (EBV), Kaposi sarcoma-associated virus (KSHV), and Helicobacter pylori, have been clearly associated with human carcinogenesis. The mucosal high-risk (HR) HPV types are the etiologic factors of cervical cancers and subset of oropharyngeal cancers. In addition, ongoing studies concerning a subgroup of HPV types that infect the skin suggest their involvement, together with ultraviolet radiation (or solar exposure), in the development of nonmelanoma skin cancer (NMSC). Biologic studies have demonstrated that the products of two early genes from the HR HPV types, E6 and E7, play a key role in cancer development. Both viral oncoproteins are able to target several cellular pathways leading to the evasion of the immune surveillance and cellular transformation. These studies also substantially contributed to our understanding of key mechanisms involved in the normal life of the cell. In the last few years, we have performed additional studies on cutaneous and mucosal HPV types and have characterized novel oncogenic viral mechanisms involved in the evasion of the immune response and/or in cellular transformation. A few examples will be presented. Citation Format: Massimo Tommasino. Role of human papillomaviruses in carcinogenesis [abstract]. In: Proceedings of the AACR Special Conference on the Microbiome, Viruses, and Cancer; 2020 Feb 21-24; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2020;80(8 Suppl):Abstract nr IA15.

A Randomized Trial of Broad Area ALA–PDT for Field Cancerization Mitigation in High-Risk Patients

The relationship between actinic keratoses (AKs) and nonmelanoma skin cancers (NMSCs) is well established. Patients with field cancerization are at high risk of developing new lesions. A treatment to interrupt new lesion formation or progression is required. To evaluate occurrence of AKs in high-risk patients after field aminolevulinic acid–photodynamic therapy (ALA–PDT). In this randomized, parallel-group, evaluator-blinded, 52-week study, patients with 4–15 facial AKs (N = 166) were random-ized (ALA 2x vs ALA 3x vs vehicle [VEH]-pooled [VEH 2x+VEH 3x], 1:1:1) to receive 2 or 3 PDT treatments (1-hour incubation) following cryotherapy at screening. More ALA-treated patients than VEH-treated patients had no AKs at week 52 (ALA 2x, 36.0%, P=0.0102; ALA 3x, 37.5%, P=0.0089; VEH, 18.9%). Week 52 lesion recurrence rates were 7.7% (P=0.0004) and 6.1% (P<0.0001) for ALA 2x and ALA 3x, respec-tively, versus 15.5% for VEH. Therapy was well tolerated; no patient requested early termination of light treatment. ALA 3x reduced NMSC development versus VEH (5 vs 12 lesions, P=0.0014). 2 or 3 ALA–PDT treatments with 1-hour incubation can significantly reduce occurrence of AKs after 1 year in patients at high risk of NMSC versus VEH–PDT (NCT02239679). J Drugs Dermatol. 2020;19(5):452-458. doi:10.36849/JDD.2020.4930.

Non-Melanoma Skin Cancer in Outdoor Workers: A Study on Actinic Keratosis in Italian Navy Personnel.

Occupational exposure to ultraviolet radiation is one of the main risk factors for non-melanoma skin cancer (NMSC) development. The most common variants of NMSC are basal cell carcinomas, squamous cell carcinomas, and actinic keratosis (AK). The latter is nowadays considered by most authors as an early squamous cell carcinoma rather than a precancerous lesion. Outdoor workers have a higher risk of developing NMSC because they spend most of the working day outside. The aim of this descriptive study was to assess the prevalence of skin lesions, especially AK, in a professional category of individuals exposed to ultraviolet (UV) radiation: the Italian Navy. From January to June 2016, a questionnaire and a total skin examination of 921 military personnel were administered by medical specialists (dermatologists) in seven different Italian Navy centres. AK was detected in 217 of 921 (23.5%) workers. Older age, outdoor occupation, longer working life, and fair skin seem to promote the development of AK. Of the 217 workers with AK, 187 (86.2%) had lesions in chronically sun-exposed skin areas. Italian Navy personnel have a high AK prevalence. Further studies are needed to investigate occupational hazards and their health effects among outdoor workers to promote protective behaviour and raise awareness of skin cancer.

A panel of criteria for comprehensive assessment of severity of ultraviolet B radiation-induced non-melanoma skin cancers in SKH-1 mice

The study of causes and cures for ultraviolet B radiation (UVB)-induced non-melanoma skin cancers (NMSC) has been greatly facilitated by use of the albino SKH-1 hairless mice. These mice develop multiple tumors of different sizes and the severity of cancer is often measured by one or more of the four criteria, namely the prevalence, multiplicity, area and volume of tumors. However, there are inherent limitations of each criterion: the prevalence and number do not account for size differences among tumors, area measurement ignores the tumor height, and volume measurement overcompensates for the height at the cost of planar dimensions. Here, using our dataset from an ongoing NMSC study, we discuss the limitations of these four criteria, and suggest refinements in measuring prevalence. We recommend the use of three more criteria, namely the Knud Thomsen tridimensional surface that apportions optimal weightage to three tumor dimensions, weekly occurrence of new tumors and tumor growth-rate to reveal initiation and growth of tumors in early and late phase of NMSC development, respectively. Together, use of this comprehensive panel of seven criteria can provide an accurate assessment of severity of NMSC and lead to a testable hypothesis whether the experimental manipulation of mice has affected the early initiation or growth phase of NMSC tumors.

NLRP1 and NLRP3 inflammasomes as a new approach to skin carcinogenesis.

Inflammasomes are key innate immune system receptors that detect pathogenic endo- and exogenous stressors like microorganisms or ultraviolet radiation (UVR) which activate the highly proinflammatory cytokines interleukin-1β and interleukin-18. Inflammasomes are not only involved in inflammation, but also in carcinogenesis and tumor progression. Due to the dynamic increase in non-melanoma skin cancers (NMSC), it has become necessary to determine how UVR, which plays a key role in NMSC development, can regulate the structure and function of inflammasomes. In the present study, the regulatory mechanisms of NOD-Like Receptor Family Pyrin Domain Containing 1 and 3 inflammasome activation as well as an effective inflammasome-mediated immune response after UVR exposition are discussed. The differences and similarities between these molecular complexes that monitor cellular health, inflammation, and skin carcinogenesis are also highlighted. Despite numerous scientific data, more studies are still required to better understand the biology of inflammasomes in skin cancer development and to explore their therapeutic potential.

The Immune System and Pathogenesis of Melanoma and Non-melanoma Skin Cancer.

Tumor development is the result of genetic derangement and the inability to prevent unfettered proliferation. Genetic derangements leading to tumorigenesis are variable, but the immune system plays a critical role in tumor development, prevention, and production. In this chapter, we will discuss the importance of the immune system as it relates to the development of skin cancer-both melanoma and non-melanoma skin cancers (NMSC).

Hydrochlorothiazide use and the risk of skin cancer in patients with hypertensive disorder: a nationwide retrospective cohort study from Korea.

Background/AimsRecent studies have shown a cumulative dose-dependent association between the use of hydrochlorothiazide (HCTZ) and skin cancer, including melanoma and non-melanoma skin cancer (NMSC) in Western Europe. However, whether this pattern is present in East Asia countries is unknown. The aim of this study was to examine the association between HCTZ use and the risk of skin cancer among a cohort of hypertensive patients in South Korea.MethodsThis was a retrospective, population-based cohort study using the database from the Health Insurance Review and Assessment Service between January 1, 2007 and June 30, 2017 in South Korea. We identified patients diagnosed with melanoma and NMSC in the cohorts of essential hypertensive patients who were treated with HCTZ or antihypertensive agents other than HCTZ. Using Cox proportional hazards regression models, the hazard ratios (HR) for skin cancer associated with HCTZ users were calculated.ResultsThe risk of melanoma was significantly lower in HCTZ-users compared with non-HCTZ users (HR, 0.85; 95% confidence interval [CI], 0.75 to 0.97; p = 0.016), and the risk of NMSC was lower in the HCTZ users but no statistically significant association was seen (HR, 0.96; 95% CI, 0.91 to 1.02; p = 0.236). High cumulative doses (≥ 50,000 mg) of HCTZ were associated with decreased risk of both NMSC (HR, 0.20; 95% CI, 0.10 to 0.38; p < 0.001) and melanoma (HR, 0.18; 95% CI, 0.04 to 0.70; p = 0.001), respectively.ConclusionsHigh cumulative use of HCTZ may have a chemopreventive effect against the development of melanoma and NMSC with clear cumulative dose-response and duration-response relationships in South Korea.

Trends in Tanning Bed Use, Motivation, and Risk Awareness in Germany: Findings from Four Waves of the National Cancer Aid Monitoring (NCAM).

Indoor tanning is an important risk factor for the development of melanoma and non-melanoma skin cancer. With our nationally representative monitoring, we aimed at describing tanning bed use, user characteristics, reasons for use, and risk awareness over time. In the framework of the National Cancer Aid Monitoring (NCAM), we collected representative data on 12,000 individuals aged 14 to 45 years in annual waves of n = 3,000 participants in Germany between 2015 and 2018. We used descriptive statistics and chi²-tests to uncover group differences. To compare data from the different waves, we calculated confidence intervals. The use of tanning beds decreased from 2015 (11.0%, 95%-CI: 9.9%–12.1%) to 2018 (8.8%, 95%-CI: 7.8%–9.8%). However, this decrease did not affect all subgroups. For instance, there was an (non-significant) increase in minors and the prevalence remained stable for individuals with immigrant background and males. Attractiveness was an important reason for tanning bed use in each wave. Over time, there was an increase in medical-related reasons for use. Furthermore, monitoring showed a decrease in risk awareness regarding tanning bed use and ultraviolet (UV) radiation. While it is a positive development that the overall use of tanning beds in Germany has decreased over time, the increasing use by minors despite the legal ban is alarming. Due to the declining risk awareness it is necessary to implement prevention and education campaigns specifically targeted at this group.

Sam68 is required for the growth and survival of nonmelanoma skin cancer.

Although targeting DNA repair signaling pathways has emerged as a promising therapeutic for skin cancer, the relevance of DNA damage responses (DDR) in the development and survival of nonmelanoma skin cancer (NMSC), the most common type of skin cancer, remains obscure. Here, we report that Src‐associated substrate during mitosis of 68 kDa (Sam68), an early signaling molecule in DDR, is elevated in skin tumor tissues derived from NMSC patients and skin lesions from Gli2‐transgenic mice. Downregulation of Sam68 impacts the growth and survival of human tumor keratinocytes and genetic ablation of Sam68 delays the onset of basal cell carcinomas (BCC) in Gli2‐transgenic mice. Moreover, Sam68 plays a critical role in DNA damage‐induced DNA repair and nuclear factor kappa B (NF‐κB) signaling pathways in keratinocytes, hence conferring keratinocyte sensitivity to DNA damaging agents. Together, our data reveal a novel function of Sam68 in regulating DDR in keratinocytes that is crucial for the growth and survival of NMSC.

LB1068 Factors relating to the development of non-melanoma skin cancers

LB1068 Factors relating to the development of non-melanoma skin cancers