Differential effects of long- and short-term exposure to PM2.5 on accelerating telomere shortening: from in vitro to epidemiological studies.

Exposure to air pollutants has been associated with DNA damage and increases the risks of respiratory diseases, such as asthma and COPD; however short- and long-term effects of air pollutants on telomere dysfunction remain unclear. We investigated the impact of short- and long-term exposure to fine particulate matter with an aerodynamic diameter below 2.5 µm (PM2.5) on telomere length in human bronchial epithelial BEAS-2B cells, and assessed the potential correlation between PM2.5 exposure and telomere length in the LIGHTS childhood cohort study. We observed that long-term, but not short-term, PM2.5 exposure was significantly associated with telomere shortening, along with the downregulation of human telomerase reverse transcriptase (hTERT) mRNA and protein levels. Moreover, long-term exposure to PM2.5 induced proinflammatory cytokine secretion, notably interleukin 6 (IL-6) and IL-8, triggered subG1 cell cycle arrest, and ultimately caused cell death. Long-term exposure to PM2.5 upregulated the LC3-II/ LC3-I ratio but led to p62 protein accumulation in BEAS-2B cells, suggesting a blockade of autophagic flux. Moreover, consistent with our in vitro findings, our epidemiological study found significant association between annual average exposure to higher PM2.5 and shortening of leukocyte telomere length in children. However, no significant association between 7-day short-term exposure to PM2.5 and leukocyte telomere length was observed in children. By combining in vitro experimental and epidemiological studies, our findings provide supportive evidence linking potential regulatory mechanisms to population level with respect to long-term PM2.5 exposure to telomere shortening in humans.

Sleep problems during early and late infancy: Diverse impacts on child development trajectories across multiple domains.

OBJECTIVE: Child developmental rate holds predictive value for early-stage developmental trajectories, yet few studies explored how sleep problems during different infancy stages impact this rate. This study aims to investigate the correlation between sleep problems and child developmental trajectories. METHODS: This study utilized a prospective national cohort of 5006 children in Taiwan. The developmental inventories covering motor, cognitive, language, and socioemotional domains were collected through questionnaire-based in-person home interviews conducted at 3, 12, 24, and 36 months. Sleep problems data, encompassing bedtime regularity, sleep duration, and sleep quality, were collected at 3 and 12 months. Child developmental rate was assessed by analyzing the slope of developmental ability estimates over a period of time. RESULTS: Bedtime regularity and high-quality sleep at 3 and 12 months were found to be significantly associated with intercepts across all domains (estimate = -0.196∼0.233, p < 0.033). Children with high-quality sleep at 3 months showed enhanced developmental slopes in socioemotional domains (estimate = 0.032, p < 0.001). Atypical sleep duration at 3 and 12 months had differential detrimental association with child development in various domains (estimate = -0.108∼-0.016, p < 0.048). CONCLUSION: The relationship between sleep problems and child development exhibited variability based on the timing of exposure to these issues. Early exposure to low-quality sleep was significantly related to developmental functions and socioemotional developmental rate, potentially leading to increased developmental disparities as children age. Inadequate sleep duration in late infancy and excessive sleep duration in early infancy were both negatively associated with child development trajectories. Policymakers can use these findings to design targeted sleep programs for optimal child development.

Measuring productive syntactic abilities in Mandarin-speaking children in Taiwan.

PURPOSE: This study aimed to develop a fine-grained measure for evaluating syntactic abilities in Mandarin-speaking children for educational and clinical purposes as a supplement to MLU. METHOD: In total, 99 typically developing children, aged 2;0 to 5;11, living in Taipei, Taiwan, participated in this study. Spontaneous language samples were elicited in free-play situations. The first 100 intelligible utterances were coded with a newly developed scheme: the Mandarin Assessment of Productive Syntax-Revised (MAPS-R). For the examination of concurrent validity, MLU was also computed. RESULTS: Significant age-related differences were observed in both MLU and MAPS-R scores. Strong correlations were found between MLU and MAPS-R scores, confirming the validity of MAPS-R as a measure of syntactic development. MAPS-R further revealed that Mandarin-speaking children expanded noun phrases with the general classifier 'GE' very early on, followed by a locative expression. Verb expansions began with resultative complements and aspect markers. Sentences with complex predication structures, such as serial verbs/pivotal sentences, were still not widely used when the MLU value is below 4.5. CONCLUSIONS: The study showed that MAPS-R is a reliable and valid measure that can provide a rich profile of the syntactic development of Mandarin-speaking children. It can be a useful reference for speech therapists to set a baseline for developing language intervention plans and to monitor their outcome.

Differential longitudinal effects of frequent sweetened food consumption at different exposure ages on child cognitive, language, and motor development.

BACKGROUND: Evidence reveals frequent sugar consumption worsens cognition in animal models, and similar effects on child development are probable. We aimed to investigate the influence of sweetened foods (SFs) on child developmental trajectories. METHODS: The prospective cohort recruited 3-month-old children in Taiwan from 1st April 2016 to 30th June 2017. Developmental inventories including cognitive, language, and motor domains, were measured at the age of 3-,12-, 24-, and 36 months old via in-person interviews. We constructed latent growth models with covariates to estimate the influence of SFs on child development. RESULTS: Ultimately, 4782 children (50.7% boys) were included in the statistical analysis. In the cognitive domain, consumption at one year of age significantly affected the intercept, but not the linear slope and quadratic term (intercept: estimate = -0.054, p < .001); consumption at two years of age significantly affected the intercept and quadratic term (intercept: estimate = -0.08, p < .001; quadratic term: estimate = -0.093, p = .026), but not the linear slope. In the language domain, only consumption at two years of age significantly affected the intercept (estimate = -0.054, p < .001). In the motor domain, consumption at two years of age significantly affected the linear slope and quadratic term (estimate = 0.080, p = .011 and estimate = -0.082, p = .048, respectively). CONCLUSION: We found SFs exposure at different times has different negative effects on child development. Early exposure to SFs harmed children's cognitive function. Relatively late exposure to SFs not only deteriorated children's cognitive and language abilities but also decelerated developmental velocity in cognitive and motor domains.

Association of exposure to environmental vanadium and manganese with lung function among young children: A population-based study.

Exposure to environmental metals has been associated with health outcomes including respiratory health. Little is known about the impact of exposure to environmental metals on lung function among young children in general population. This study aimed to investigate the associations of exposure to metals with lung function among young children in a population-based cohort. A total of 1488 children aged 5-8 years attended a follow-up visit as part of the Longitudinal Investigation of Global Health in Taiwanese Schoolchildren (LIGHTS) cohort. We measured urinary samples of vanadium (median: 1.21 ng/mL; interquartile range (IQR): 0.73-1.98), manganese (median: 0.23 ng/mL; IQR: 0.13-0.47), arsenic (median: 40.51 ng/mL; IQR: 21.66-70.49), nickel (median: 1.09 ng/mL; IQR: 0.31-3.60), and cadmium (median: 0.26 ng/mL; IQR: 0.11-0.43) and performed lung function tests. Urinary vanadium concentrations were inversely associated with FVC (ß coefficient for the highest quartile versus the other quartiles: -33.40, p = 0.001), FEV1 (ß: -41.31, p < 0.001), FEV1/FVC ratio (ß: -1.00, p = 0.009), PEF (ß: -92.12, p = 0.004), and FEF25-75 (ß: -82.85, p < 0.001), after adjusting for relevant confounders. Urinary manganese concentrations were inversely associated with FVC (ß: -26.60, p = 0.007), FEV1 (ß: -31.62, p = 0.001), PEF (ß: -84.86, p = 0.009), and FEF25-75 (ß: -69.21, p = 0.002). Stratification analyses found inverse associations of urinary vanadium and manganese concentrations with lung function parameters predominantly among children exposed to environmental tobacco smoke. We did not find significant associations of urinary arsenic, nickel, and cadmium concentrations with lung function parameters. In conclusion, this study adds new evidence showing inverse associations of vanadium and manganese exposure with lung function among young children in the general population. Children with environmental tobacco smoke exposure are particularly vulnerable to adverse impact of vanadium and manganese exposure on lung function.

Developing early lexical composition in Mandarin-speaking children: A longitudinal study.

This study investigated the developmental pattern of early lexical production and composition in Mandarin-speaking children. Forty Mandarin-speaking children and their parents participated in this one-and-a-half-year longitudinal study, and naturalistic samples of parent-to-child speech in toy play were collected when the children were 1;8, 2;2, and 3;0. The results showed that children's lexical production increased significantly between ages 1;8 and 3;0. The proportion of closed-class words increased significantly with age, whereas the proportion of common nouns showed the inverse pattern, indicating the role of grammatical words increased as the children grew. Furthermore, nouns and verbs were predominant in Mandarin-speaking children between ages 1;8 and 3;0, and Mandarin-speaking children used more verbs than nouns at 2;2 and 3;0 in the toy play context. The longitudinal study clarifies early lexical development in Mandarin-speaking children, which provides a valuable contrast for different language systems.

Influence of parental play on Taiwanese 3-year-olds' executive function: Through the path of motor skills.

Parenting is known to impact children's executive function (EF) skills. However, nearly all the evidence comes from analyses of mother-child interaction. Using the National Longitudinal Study of Child Development and Care Database in Taiwan, the relations between both mother-child and father-child interaction and 3-year-olds' EF were investigated in 2,164 families. The results showed that mothers interacted with their children differently from fathers in terms of time distribution. Mothers were more equally involved in all aspects of parental involvement, whereas fathers spent more time in play. In addition, both mother-child and father-child play contributed to children's EF; however, the mediating effect of child motor skills was more prominent for father-child play. This study not only suggests a potential distinct and complementary role of fathers in young children's EF development but also indicates a unique mediating effect of motor skills in the path from parent-child play to child EF. Implications for parent education are discussed. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Protective mechanism of ferulic acid against neomycin-induced ototoxicity in zebrafish.

Ototoxicity refers to damage of sensory hair cells and functional hearing impairment following aminoglycosides exposure. Previously, we have determined that ferulic acid (FA) protected hair cells against serial concentrations of neomycin-induced ototoxic damage. The aim of the present study is to assess the mechanism and effects of FA on neomycin-induced hair cells loss and impact on mechanosensory-mediated behaviors alteration using transgenic zebrafish (pvalb3b: TagGFP). We first identified the optimal protective condition as pre/co-treatment method in early fish development. Pretreatment of the larvae with FA significantly protected against neomycin-induced hair cells loss through preventing neomycin passed through the cytoplasm of hair cells, and subsequently decreased reactive oxygen species production and TUNEL signals in 4 day post-fertilization (dpf) transgenic zebrafish larvae. Moreover, preservation of functional hair cells correlated directly with rescue of the altered swimming behavior, indicates FA pretreatment protects against neomycin ototoxic damage in 7-dpf transgenic zebrafish larvae. Together, our findings unravel the otoprotective role of FA as an effective agent against neomycin-induced ototoxic effects and offering the theoretical foundation for discovering novel candidates for hearing protection.

The performance and predictors of Chinese character writing in children with attention deficit hyperactivity disorder.

Writing difficulties are common in children with attention deficit hyperactivity disorder (ADHD). Preliminary evidence suggests that early character writing ability is fundamental for later writing composition and academic achievement. Critical factors of different character writing tasks in children with ADHD, however, remain unclear. This study aims to describe the performance and identify predictors of Chinese character writing in children with ADHD. Thirty Mandarin Chinese-speaking children with ADHD (7.16 ± 0.59 years) and thirty matched peers (7.21 ± 0.57 years) were recruited from northern Taiwan. They were evaluated with the Battery of Chinese Basic Literacy (BCBL); the Chinese version of the Test of Nonverbal Intelligence, fourth edition (C-TONI-4); the Chinese version of the Peabody Picture Vocabulary Test-Revised (C-PPVT-R); orthographic awareness test; character naming test; and the Bruininks Oseretsky Test of Motor Proficiency, second edition (BOT-2). The results showed that Mandarin Chinese-speaking children with ADHD scored lower than their typically developing peers on both dictation and copying subtests. After controlling for age, orthographic awareness and inattention were identified as important predictors of character dictation; while, manual dexterity was a critical predictor of character copying in children with ADHD. The results demonstrated that character writing problems may exist in Mandarin Chinese-speaking children with ADHD, and cognitive-linguistic and fine-motor skills have varying contributions to Chinese character writing tasks.

Differential Trajectories of Fathers' Postpartum Depressed Mood: A Latent Class Growth Analysis Approach.

Parental psychological well-being is essential to the wellness of the family. However, longitudinal investigations into fathers' postpartum depressed mood are limited. This study aimed to identify the typologies of depressed mood trajectories over the first year postpartum among Taiwanese fathers and to examine the factors associated with such typologies. We retrieved data from a nationwide longitudinal study on child development and care in Taiwan. A total of 396 fathers, who completed at least one of the three interviews when their children were 3, 6, and 12 months old between 2016 and 2017, were included in this analysis. Conditional latent class growth analysis was conducted to identify the classifications of the fathers' depressed mood trajectories in the first year postpartum and to estimate the effects of covariates on individuals' membership of a trajectory class. Three classes of depressed mood trajectories were identified. The high increasing group consisted of 11% of the participants; the moderate increasing and the low decreasing groups consisted of 28% and 61% of the participants, respectively. Financial stress was associated with the fathers' likelihood of being in the high increasing group compared with their likelihood of being in the low decreasing group (OR = 2.28, CI = 1.16-4.47). The result may be related to the difference in gender roles and social expectations.

Depressed mood trajectories in the first year postpartum among Taiwanese mothers: Associations with perceived support, financial stress, and marital satisfaction.

This study examined the typology of depressed mood trajectories and the associated factors over the first year postpartum among Taiwanese mothers. Data of 4332 mothers from a nationwide longitudinal study on child development and care were analyzed. Three classes of depressed mood trajectories were identified, two with lower initial scores and a decreasing trajectory and one with a higher initial score and an increasing trajectory. Subjective financial stress, perceived support, and marital satisfaction were significant in predicting mothers' membership of the depressed mood trajectory classes. The results highlighted the individual susceptibility to the postpartum depressed mood among Taiwanese mothers.

Particulate matter causes telomere shortening and increase in cellular senescence markers in human lung epithelial cells.

Exposure to particulate matter (PM) has been associated with DNA damage, but the relationships between PM, telomere length and cellular senescence remain unclear. This study aimed to investigate the effects and potential mechanisms of PM on telomere length and cellular senescence in human lung epithelial cells. Human lung epithelial A549 cells were exposed to PM for 24 h. Cell viability and cytotoxicity were measured by the WST-1 assay and the lactate dehydrogenase release, respectively. Cellular uptake of PM was observed using transmission electron microscopy. Telomere length was measured using qPCR and expressed as T/S ratio. Cell cycle progression was analyzed by flow cytometry. Expression of human telomerase reverse transcriptase (hTERT) and cell cycle regulators was measured using mRNA by qPCR and protein levels by Western blot. Cellular senescence was determined by the expression of senescence-associated ß-galactosidase (SA-ß-Gal) with fluorescent microscopy and flow cytometry. Exposed to PM at the concentration of 200 µg/ml decreased cell viability and increased LDH levels in culture medium. Remarkably increased uptake of PM, shortening of telomere length, induction of G0/G1 phase arrest, and increased expression of senescence hallmarks were observed after exposure to PM in A549 cells. PM exposure induced upregulation of p21 and downregulation of proliferating cell nuclear antigen (PCNA) and hTERT expression, but no significant change in p53 expression, in A549 cells. Overall, exposure to PM may downregulate hTERT and PCNA through p53-independent induction of p21 expression, leading to telomere shortening, G0/G1 arrest and the onset of cellular senescence in human lung epithelial cells.

Metabolomic differences of exhaled breath condensate among children with and without asthma.

BACKGROUND: There remains an unmet need in objective tests for diagnosing asthma in children. The objective of this study was to investigate the potential of metabolomic profiles of exhaled breath condensate (EBC) to discriminate stable asthma in Asian children in the community. METHODS: One hundred and sixty-five Asian children (92 stable asthma and 73 non-asthmatic controls) participating in a population-based cohort were enrolled and divided into training and validation sets. Nuclear magnetic resonance-based metabolomic profiles of EBC samples were analyzed by using orthogonal partial least squares discriminant analysis. RESULTS: EBC metabolomic signature (lactate, formate, butyrate, and isobutyrate) had an area under the receiver operator characteristic curve (AUC) of 0.826 in discriminating children with and without asthma in the training set, which significantly outperformed FeNO (AUC = 0.574; P < .001) and FEV1 /FVC % predicted (AUC = 0.569; P < .001). The AUC for EBC metabolomic signature was 0.745 in the validation set, which was slightly but not significantly lower than in the testing set (P = .282). We further extrapolated two potentially involved metabolic pathways, including pyruvate (P = 1.67 × 10-3 ; impact: 0.14) and methane (P = 1.89 × 10-3 ; impact: 0.15), as the most likely divergent metabolisms between children with and without asthma. CONCLUSION: This study provided evidence supporting the role of EBC metabolomic signature to discriminate stable asthma in Asian children in the community, with a discriminative property outperforming conventional clinical tests such as FeNO or spirometry.

Association between maternal age at delivery and allergic rhinitis in schoolchildren: A population-based study.

BACKGROUND: Some studies have shown associations of maternal age at delivery with asthma and food allergy in offspring. However, the relationship between maternal age at delivery and allergic rhinitis is largely unclear. This study aimed to investigate the association between maternal age at delivery and allergic rhinitis in a population sample of Asian children, and to explore potential effect modifiers. METHODS: A total of 1344 singleton-birth children (763 boys, 56.8%; mean age, 6.4 years) participating in the Longitudinal Investigation of Global Health in Taiwanese Schoolchildren (LIGHTS) cohort were evaluated by a modified International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire and interviewed by pediatricians. Allergic sensitization was determined by using Phadiatop Infant. Multiple logistic regression models with covariates adjustment were performed to investigate the association of maternal age at delivery with allergic rhinitis and allergic sensitization in offspring. RESULTS: Among 1344 study children, 793 (59%) had physician-diagnosed allergic rhinitis. Advanced maternal age at delivery (≥40 years) was significantly associated with increased odds of allergic rhinitis (adjusted odds ratio [AOR] = 4.58, 95% confidence interval [CI]: 1.90-11.03) and allergic sensitization (AOR = 2.86, 95% CI: 1.13-7.22) in offspring. A sex-stratified analysis revealed that the association of advanced maternal age with allergic rhinitis was statistically significant only in female offspring (AOR = 7.02, 95% CI: 1.89-26.14). Stratified analyses by birth order or environmental tobacco smoke exposure during pregnancy did not reveal any significant differences. CONCLUSION: Advanced maternal age at delivery was associated with increased risk of allergic rhinitis in Asian children, probably more pronounced among girls.

A longitudinal study of maternal interaction strategies during joint book-reading in Taiwan.

This longitudinal study examines change in maternal interaction strategies in Taiwanese mothers across time, and the synchronic and diachronic relationships between maternal interaction strategies and children's language and early literacy skills. Forty-two mother-child dyads participated in this study. Their interactions during joint book-reading were tape-recorded, transcribed, and analyzed when the children were fourteen, twenty-six, and thirty-six months of age. The children received a battery of language and early literacy tests when they were thirty-six months old. Findings showed that Taiwanese mothers adjusted their use of interaction strategies as their children grew. Maternal use of description, performance, prediction inference, and print-related talk were positively correlated with their children's language and literacy skills. Significant negative correlations were found between use of task-behavioral regulation strategy and text reading in mothers and their children's language performance. This study suggests that age-appropriate interaction strategies are important for children's language and early literacy development.

Childhood otitis media: Relationship with daycare attendance, harsh parenting, and maternal mental health.

Psychological stress has been linked to developmental problems and poor health in children, but it is unclear whether it is also related to otitis media (OM). As part of a long-term study surveying the characteristics of childcare and development in Taiwan, we analyzed the relationship between OM and sources of psychological stress in children, such as poor maternal mental health and harsh parental discipline. We analyzed the data of 1998 children from the "Kids in Taiwan: National Longitudinal Study of Child Development & Care (KIT) Project" at the age of 3 years. Using bivariate and multivariate logistic regression models, we tested several risk factors as potential independent predictors of two outcomes: parent-reported incidence of OM and child health. The proportion of children who had developed OM in the first 3 years of their life was 12.5%. Daycare attendance (odds ratio [OR]: 1.475; 95% confidence interval [CI]: 1.063-2.046), poor maternal mental health (OR: 1.913; 95% CI: 1.315-2.784), and harsh parental discipline (OR: 1.091; 95% CI: 1.025-1.161) correlated with parent-reported occurrence of OM. These findings suggest that providing psychosocial support to both parents and children might be a novel strategy for preventing OM.

Hsp90α Mediates BMI1 Expression in Breast Cancer Stem/Progenitor Cells through Facilitating Nuclear Translocation of c-Myc and EZH2.

Heat shock protein 90 (Hsp90) is a molecular chaperone that facilitates the correct folding and functionality of its client protein. Numerous Hsp90-client proteins are involved in cancer development. Thus, Hsp90 inhibitors have potential applications as anti-cancer drugs. We previously discovered that Hsp90α expression increased in breast cancer stem cells (BCSCs), which can initiate tumorigenesis and metastasis and resist treatment. In the present study, we further demonstrated that 17-dimethylaminoethylamino-17-demethoxygeldanamycin (17-DMAG), an inhibitor of Hsp90, could suppress the self-renewal of BCSCs by downregulating B lymphoma Mo-MLV insertion region 1 homolog (BMI1), a polycomb family member with oncogenic activity in breast cancer. Through immunoprecipitation analysis, we found that BMI1 did not interact with Hsp90α and that the downregulation of BMI1 by 17-DMAG was mediated by the inhibition of c-Myc and enhancement of zeste homolog 2 (EZH2) expression. The transcriptional and BMI1 promoter-binding activities of c-Myc in BCSCs were inhibited by 17-DMAG treatment. The overexpression of EZH2 attenuated the inhibitory effect of 17-DMAG on BMI1 and c-Myc expression. Furthermore, Hsp90α could be co-immunoprecipitated with c-Myc and EZH2 and bind to the BMI1 promoter. Treatment with 17-DMAG decreased the nuclear expression of EZH2 and c-Myc but not that of Hsp90α. In conclusion, our data suggested that Hsp90α could positively regulate the self-renewal of BCSCs by facilitating the nuclear translocation of c-Myc and EZH2 to maintain BMI1 expression.

Ovatodiolide Inhibits Breast Cancer Stem/Progenitor Cells through SMURF2-Mediated Downregulation of Hsp27.

Cancer stem/progenitor cells (CSCs) are a subpopulation of cancer cells involved in tumor initiation, resistance to therapy and metastasis. Targeting CSCs has been considered as the key for successful cancer therapy. Ovatodiolide (Ova) is a macrocyclic diterpenoid compound isolated from Anisomeles indica (L.) Kuntze with anti-cancer activity. Here we used two human breast cancer cell lines (AS-B145 and BT-474) to examine the effect of Ova on breast CSCs. We first discovered that Ova displayed an anti-proliferation activity in these two breast cancer cells. Ova also inhibited the self-renewal capability of breast CSCs (BCSCs) which was determined by mammosphere assay. Ova dose-dependently downregulated the expression of stemness genes, octamer-binding transcription factor 4 (Oct4) and Nanog, as well as heat shock protein 27 (Hsp27), but upregulated SMAD ubiquitin regulatory factor 2 (SMURF2) in mammosphere cells derived from AS-B145 or BT-474. Overexpression of Hsp27 or knockdown of SMURF2 in AS-B145 cells diminished the therapeutic effect of ovatodiolide in the suppression of mammosphere formation. In summary, our data reveal that Ova displays an anti-CSC activity through SMURF2-mediated downregulation of Hsp27. Ova could be further developed as an anti-CSC agent in the treatment of breast cancer.

Advances and Applications of Ion Torrent Personal Genome Machine in Cutaneous Squamous Cell Carcinoma Reveal Novel Gene Mutations.

The Ion Torrent Personal Genome Machine (Ion PGM) is a semiconductor-based sequencing technology that is high quality, scalable, and economic. Its applications include genomic sequencing, drug resistance testing, microbial characterization, and targeted sequencing in cancer studies. However, little is known about the application of Ion PGM in cutaneous squamous cell carcinoma (cSCC). We therefore investigated the utility and validity of Ion PGM in cSCC and also gained a better understanding of the underlying molecular biology of cSCC. We detected novel gene mutations (KDR, FGFR2, and EGFR) in two cSCC patients. Moreover, we validated these mutations by pyrosequencing and Sanger sequencing. Our results indicated that the mutation screen using Ion PGM is consistent with traditional sequencing methods. Notably, these identified mutations were present at significantly higher rates in high-risk cSCC. Our results demonstrate a method to detect targetable genes in high-risk cSCC, and suggest that Ion PGM may enable therapeutic decision-making and future potential targets for personalized therapies in cSCC.