Loss of fragile WWOX gene leads to senescence escape and genome instability.

Induction of DNA damage response (DDR) to ensure accurate duplication of genetic information is crucial for maintaining genome integrity during DNA replication. Cellular senescence is a DDR mechanism that prevents the proliferation of cells with damaged DNA to avoid mitotic anomalies and inheritance of the damage over cell generations. Human WWOX gene resides within a common fragile site FRA16D that is preferentially prone to form breaks on metaphase chromosome upon replication stress. We report here that primary Wwox knockout (Wwox-/-) mouse embryonic fibroblasts (MEFs) and WWOX-knockdown human dermal fibroblasts failed to undergo replication-induced cellular senescence after multiple passages in vitro. Strikingly, by greater than 20 passages, accelerated cell cycle progression and increased apoptosis occurred in these late-passage Wwox-/- MEFs. These cells exhibited γH2AX upregulation and microsatellite instability, indicating massive accumulation of nuclear DNA lesions. Ultraviolet radiation-induced premature senescence was also blocked by WWOX knockdown in human HEK293T cells. Mechanistically, overproduction of cytosolic reactive oxygen species caused p16Ink4a promoter hypermethylation, aberrant p53/p21Cip1/Waf1 signaling axis and accelerated p27Kip1 protein degradation, thereby leading to the failure of senescence induction in Wwox-deficient cells after serial passage in culture. We determined that significantly reduced protein stability or loss-of-function A135P/V213G mutations in the DNA-binding domain of p53 caused defective induction of p21Cip1/Waf1 in late-passage Wwox-/- MEFs. Treatment of N-acetyl-L-cysteine prevented downregulation of cyclin-dependent kinase inhibitors and induced senescence in Wwox-/- MEFs. Our findings support an important role for fragile WWOX gene in inducing cellular senescence for maintaining genome integrity during DDR through alleviating oxidative stress.

Collaboration between Trinuclear Aluminum Complexes Bearing Bipyrazoles in the Ring-Opening Polymerization of ε-Caprolactone.

Trinuclear aluminum complexes bearing bipyrazoles were synthesized, and their catalytic activity for ε-caprolactone (CL) polymerization was investigated. DBu2Al3Me5 exhibited higher catalytic activity than did the dinuclear aluminum complex LBu2Al2Me4 (16 times as high for CL polymerization; [CL]:[DBu2Al3Me5]:[BnOH] = 100:0.5:5, [DBu2Al3Me5] = 10 mM, conversion 93% after 18 min at room temperature). Density functional theory calculations revealed a polymerization mechanism in which CL first approached the central Al atom and then moved to an external Al. The coordinated CL ring was opened because the repulsion of two tert-butyl groups on the ligands pushed an alkoxide initiator on an external Al to initiate CL. In these trinuclear Al catalysts, the central Al plays a role in monomer capture and then collaborates with the external Al to activate CL, accelerating polymerization.

RISK OF RETINAL VEIN OCCLUSION IN PATIENTS WITH PSORIASIS: A Population-Based Cohort Study.

PURPOSE: To investigate the risk of developing retinal vein occlusion (RVO) in patients with psoriasis. METHODS: In this retrospective population-based cohort study, 30,198 patients with psoriasis (Psoriasis((+)) group) and 30,198 controls without psoriasis (Psoriasis((−)) group) between 2001 and 2006 from the Taiwan National Health Insurance Research Database were selected. RESULTS: The incidence of RVO was 1.46 times higher in the Psoriasis((+)) group than in the Psoriasis((−)) group (3.61 vs. 2.47/10,000 person-years) (adjusted hazard ratio = 1.50; 95% confidence interval = 1.07-2.10) calculated using Cox proportional hazard regression. Age was an independent risk factor for RVO (adjusted hazard ratio: 11.9 for patients 65 years or older vs. 1.00 for those 0-49 years old). In the 65 years or older Psoriasis((+)) group, the incidence of developing RVO was 1.97 times higher (95% confidence interval = 1.19-3.26) than in the 65 years or older Psoriasis((−)) group. In Psoriasis((+)) women, the incidence of developing RVO was 1.82 times higher (95% = 1.05-3.14) than in Psoriasis((−)) women. For the subgroup with comorbid hypertension, the incidence of developing RVO was 2.07 times higher (95% confidence interval = 1.22-3.50) in the Psoriasis((+)) group than in the Psoriasis((−)) group. CONCLUSION: Psoriasis was significantly associated with a higher risk of developing RVO.

Benzyl butyl phthalate induces migration, invasion, and angiogenesis of Huh7 hepatocellular carcinoma cells through nongenomic AhR/G-protein signaling.

BACKGROUND: The widespread use of phthalates as plasticizers has raised public health concerns regarding their adverse effects, including an association with cancer. Although animal investigations have suggested an association between phthalate exposure and hepatocellular carcinoma, the mechanisms are unknown. METHODS: The hepatocellular carcinoma cell line Huh7 was treated with benzyl butyl phthalate (BBP), and then analyzed by total internal reflection fluorescence microscopy, confocal microscopy and double immunogold transmission electron microscopy. Following BBP treatment, mRNA levels were measured by RT-PCR, protein levels were measured using western blot, and vascular endothelial growth factor levels were measured by an enzyme-linked immunosorbent assay. Cell migration and invasion assays were evaluated by transwell, and angiogenesis were performed by a tube formation assay. Nude mice were used to investigate metastasis and angiogenesis in vivo. RESULTS: BBP affected hepatocellular carcinoma progression through the aryl hydrocarbon receptor (AhR) and that benzyl butyl phthalate (BBP) stimulated AhR at the cell surface, which then interacted with G proteins and triggered a downstream signaling cascade. BBP activated AhR through a nongenomic action involving G-protein signaling rather than the classical genomic AhR action. BBP treatment promoted cell migration and invasion in vitro and metastasis in vivo via the AhR/Gß/PI3K/Akt/NF-κB pathway. In addition, BBP induced both in vitro and in vivo angiogenesis through the AhR/ERK/VEGF pathway. CONCLUSIONS: These findings suggest a novel nongenomic AhR mechanism involving G-protein signaling induced by phthalates, which contributes to tumor progression of hepatocellular carcinoma.

A modern approach with follower-leading clustering algorithm for visualizing author collaborations and article themes in skin cancer research: A bibliometric analysis.

BACKGROUND: Skin cancers (SCs) arise due to the proliferation of atypical cells that have the potential to infiltrate or metastasize to different areas of the body. There is a lack of understanding regarding the country-based collaborations among authors (CBCA) and article themes on SCs. A clustering algorithm capable of categorizing CBCA and article themes on skin cancer is required. This study aimed to apply a follower-leading clustering algorithm to classify CBCA and article themes and present articles that deserve reading in recent ten years. METHODS: Between 2013 and 2022, a total of 6526 articles focusing on SC were extracted from the Web of Science core collection. The descriptive, diagnostic, predictive, and prescriptive analytics model was employed to visualize the study results. Various visualizations, including 4-quadrant radar plots, line charts, scatter plots, network charts, chord diagrams, and impact beam plots, were utilized. The category, journal, authorship, and L-index score were employed to assess individual research achievements. Diagnostic analytics were used to cluster the CBCA and identify common article themes. Keyword weights were utilized to predict article citations, and noteworthy articles were highlighted in prescriptive analytics based on the 100 most highly cited articles on SC (T100SC). RESULTS: The primary entities contributing to SC research include the United States, the University of California, San Francisco in US, dermatology department, and the author Andreas Stang from Germany, who possess higher category, journal, authorship, and L-index scores. The Journal of the American Academy of Dermatology has published the highest number of articles (n = 336, accounting for 5.16% of the total). From the T100SC, 7 distinct themes were identified, with melanoma being the predominant theme (92% representation). A strong correlation was observed between the number of article citations and the keyword weights (F = 81.63; P < .0001). Two articles with the highest citation counts were recommended for reading. CONCLUSION: By applying the descriptive, diagnostic, predictive, and prescriptive analytics model, 2 noteworthy articles were identified and highlighted on an impact beam plot. These articles are considered deserving of attention and could potentially inspire further research in the field of bibliometrics, focusing on relevant topics related to melanoma.

Using temporal heatmaps to identify worthwhile articles on immune checkpoint blockade for melanoma (ICBM) in Mainland China, Hong Kong, and Taiwan since 2000: A bibliometric analysis.

BACKGROUND: Melanoma is a life-threatening form of skin cancer. Due to its remarkable effectiveness, the immune checkpoint blockade is widely used to treat melanoma (ICBM). No research has been conducted on ICBM for identifying the most readable articles. A bibliometric analysis of 100 top-cited ICBM (T100ICBM) in recent decades is required to highlight articles worth reading. METHODS: Based on the Web of Science Core Collection, we summarized the articles on ICBM published in each year from 2000 to 2022, with first authors from Mainland China, Hong Kong, and Taiwan (CHT). Using the CJAL score, data extraction and visualization of the distribution of ICBM publications were conducted on 2718, and 100 top-cited articles, respectively. We used the temporal heatmap to identify the most readable articles. Four descriptive, diagnostic, predictive, and prescriptive analytics (called DDPP model) were applied to describe the features of T100ICBM articles. The absolute advantage coefficient was used to determine the dominance extent of the most influential region, institute, department, and author. RESULTS: A total of 2718 publications was included after removing first or corresponding authors who were not affiliated with CHT. Publications by year showed a sharp increase from 2014 onward and either peaked in 2022 or have not yet peaked. It was evident that there was a large difference between the number of publications in provinces/metropolitan cities/regions on CHT. Beijing, Sichuan University, Oncology, and Guo Jun from Beijing are the most prolific and influential region, institute, department, and author. When comparing research achievements to the next productive authors based on the CJAL score, only Dr Jun has a medium effect of dominance (=0.60). On the basis of their consecutive growth in citations over the past 4 years, 20 T100ICBM articles were recommended for readers. CONCLUSION: The field of ICBM is growing rapidly, and Beijing and Sichuan University are taking the lead in CHT. Furthermore, the study provides references for worth-reading articles using the temporal heatmap. Future research hot spots may focus on these 4 themes of immunotherapy, melanoma, metastatic melanoma, regulatory T cells, cells, and activation, which may pave the way for additional study.

Web-Based Skin Cancer Assessment and Classification Using Machine Learning and Mobile Computerized Adaptive Testing in a Rasch Model: Development Study.

BACKGROUND: Web-based computerized adaptive testing (CAT) implementation of the skin cancer (SC) risk scale could substantially reduce participant burden without compromising measurement precision. However, the CAT of SC classification has not been reported in academics thus far. OBJECTIVE: We aim to build a CAT-based model using machine learning to develop an app for automatic classification of SC to help patients assess the risk at an early stage. METHODS: We extracted data from a population-based Australian cohort study of SC risk (N=43,794) using the Rasch simulation scheme. All 30 feature items were calibrated using the Rasch partial credit model. A total of 1000 cases following a normal distribution (mean 0, SD 1) based on the item and threshold difficulties were simulated using three techniques of machine learning-naïve Bayes, k-nearest neighbors, and logistic regression-to compare the model accuracy in training and testing data sets with a proportion of 70:30, where the former was used to predict the latter. We calculated the sensitivity, specificity, receiver operating characteristic curve (area under the curve [AUC]), and CIs along with the accuracy and precision across the proposed models for comparison. An app that classifies the SC risk of the respondent was developed. RESULTS: We observed that the 30-item k-nearest neighbors model yielded higher AUC values of 99% and 91% for the 700 training and 300 testing cases, respectively, than its 2 counterparts using the hold-out validation but had lower AUC values of 85% (95% CI 83%-87%) in the k-fold cross-validation and that an app that predicts SC classification for patients was successfully developed and demonstrated in this study. CONCLUSIONS: The 30-item SC prediction model, combined with the Rasch web-based CAT, is recommended for classifying SC in patients. An app we developed to help patients self-assess SC risk at an early stage is required for application in the future.

Panfolliculoma: Report of the Youngest Case and Literature Review of Its Histopathologic Variants.

Background: Panfolliculoma (PF) is a relative rare, benign follicular tumor comprised of all elements of the hair follicle, with a limited number of cases reported in the literature. Articles on the demographic and pathological analysis of this tumor are also lacking. Case presentation: In this report, we presented an unusual case of cystic PF on the back of a 14-year-old male, and we performed a thorough literature review and analysis of all previously reported cases. Conclusions: PF is a rare benign follicular neoplasm with characteristic differentiation toward all components of the hair follicle. In our analysis, PF occurred most frequently on the head region and was usually diagnosed in middle- to old-aged persons, with cystic PF being the most common histologic subtype. Since this tumor is rare and easily misdiagnosed as other tumors both clinically and pathologically, a thorough understanding of the histopathological manifestations and differential diagnosis of this tumor is necessary for both dermatologists and pathologists.

Citation analysis of the 100 top-cited articles on the topic of hidradenitis suppurativa since 2013 using Sankey diagrams: Bibliometric analysis.

BACKGROUND: Hidradenitis suppurativa (HS) is a chronic, inflammatory and debilitating dermatosis characterized by painful nodules, sinus tracts and abscesses in apocrine gland-bearing areas that predominantly affect women worldwide. New therapeutic interventions based on the clinical manifestations of patients have recently been introduced in numerous articles. However, which countries, journals, subject categories, and articles have the ultimate influence remain unknown. This study aimed to display influential entities in 100 top-cited HS-related articles (T100HS) and investigate whether medical subject headings (i.e., MeSH terms) can be used to predict article citations. METHODS: T100HS data were extracted from PubMed since 2013. Subject categories were classified by MeSH terms using social network analysis. Sankey diagrams were applied to highlight the top 10 influential entities in T100HS from the three aspects of publication, citations, and the composited score using the hT index. The difference in article citations across subject categories and the predictive power of MeSH terms on article citations in T100HS were examined using one-way analysis of variance and regression analysis. RESULTS: The top three countries (the US, Italy, and Spain) accounts for 54% of the T100HS. The T100HS impact factor (IF) is 12.49 (IF = citations/100). Most articles were published in J Am Acad Dermatol (15%; IF = 18.07). Eight subject categories were used. The "methods" was the most frequent MeSH term, followed by "surgery" and "therapeutic use". Saunte et al, from Roskilde Hospital, Denmark, had 149 citations in PubMed for the most cited articles. Sankey diagrams were used to depict the network characteristics of the T100HS. Article citations did not differ by subject category (F(7, 92) = 1.97, P = .067). MeSH terms were evident in the number of article citations predicted (F(1, 98) = 129.1106; P < .001). CONCLUSION: We achieved a breakthrough by displaying the characteristics of the T100HS network on the Sankey diagrams. MeSH terms may be used to classify articles into subject categories and predict T100HS citations. Future studies can apply the Sankey diagram to the bibliometrics of the 100 most-cited articles.

Using the Sankey diagram to visualize article features on the topics of whole-exome sequencing (WES) and whole-genome sequencing (WGS) since 2012: Bibliometric analysis.

BACKGROUND: Sequencing technologies, such as whole-exome sequencing (WES) and whole-genome sequencing (WGS), have been increasingly applied to medical research in recent years. Which countries, journals, and institutes (called entities) contributed most to the fields (WES/WGS) remains unknown. Temporal bar graphs (TBGs) are frequently used in trend analysis of publications. However, how to draw the TBG on the Sankey diagram is not well understood in bibliometrics. We thus aimed to investigate the evolution of article entities in the WES/WGS fields using publication-based TBGs and compare the individual research achievements (IRAs) among entities. METHODS: A total of 3599 abstracts downloaded from icite analysis were matched to entities, including article identity numbers, citations, publication years, journals, affiliated countries/regions of origin, and medical subject headings (MeSH terms) in PubMed on March 12, 2022. The relative citation ratio (RCR) was extracted from icite analysis to compute the hT index (denoting the IRA, taking both publications and citations into account) for each entity in the years between 2012 and 2021. Three types of visualizations were applied to display the trends of publications (e.g., choropleth maps and the enhanced TBGs) and IRAs (e.g., the flowchart on the Sankey diagram) for article entities in WES/WGS. RESULTS: We observed that the 3 countries (the US, China, and the UK) occupied most articles in the WES/WGS fields since 2012, the 3 entities (i.e., top 5 journals, research institutes, and MeSH terms) were demonstrated on the enhanced TBGs, the top 2 MeSH terms were genetics and methods in WES and WGS, and the IRAs of 6 article entities with their hT-indices were succinctly and simultaneously displayed on a single Sankey diagram that was never launched in bibliographical studies. CONCLUSION: The number of WES/WGS-related articles has dramatically increased since 2017. TBGs, particularly with hTs on the Sankey, are recommended for research on a topic (or in a discipline) to compare trends of publications and IRAs for entities in future bibliographical studies.

Diagnostic clues for differentiating Merkel cell carcinoma from lymphoma in fine-needle aspiration cytology.

Nodal fine needle aspiration (FNA) is usually the first procedure in the work-up of malignancy of unknown primary. Merkel cell carcinoma (MCC) is an aggressive cutaneous cancer more common in Caucasians but rare among Asians. It is a diagnostic challenge in evaluating FNA from a metastatic MCC without the knowledge of a current or prior history of skin cancer. We report the case of a Taiwanese male with cervical and axillary masses. The diagnosis of the FNA from the axillary lymph node was lymphoproliferative lesion suspicious for lymphoma. The histopathological evaluation of nodal biopsy revealed a metastatic neuroendocrine carcinoma and the subsequent excision of the right palm tumor confirmed MCC. Retrospective review of the FNA and imprint cytology smears of the nodal biopsy showed nuclear molding, Indian filing and rare cytoplasmic pale bodies, but no lymphoglandular bodies. Cytologically metastatic MCC may mimic small round cell tumor including lymphoma, we consider these three cytological features as additional diagnostic clues for metastatic MCC. In this report, we present the cytologic and pathological features of this metastatic MCC and discuss the differential diagnosis of the cytologic mimickers.

Predicting article citations using data from 100 top-cited publications in the field of Psoriasis Vulgaris and biological agents (PVBA) since 1991: A bibliometric analysis.

BACKGROUND: Psoriasis Vulgaris is a chronic inflammatory disease characterized by keratinocyte hyperproliferation. Bibliometric analysis helps determine the most influential article on the topic of "Psoriasis Vulgaris and biological agents (PVBAs)", and what factors affect article citation remain unclear. This study aims (1) to identify the top 100 most cited articles in PVBA (PVBA100 for short) from 1991 to 2020, (2) to visualize dominant entities on one diagram using data in PVBA100, and (3) to investigate whether medical subject headings (MeSH terms) can be used to predict article citations. METHODS: The top 100 most cited articles relevant to PVBA (1991-2020) were downloaded by searching the PubMed database. Citation analysis was applied to compare the dominant roles in article types and topic categories using pyramid plots. Social network analysis (SNA) and Sankey diagrams were applied to highlight prominent entities. We examined the MeSH prediction effect on article citations using its correlation coefficients. RESULTS: The most frequent article types and topic categories were research support by institutes (46%) and drug therapy (88%), respectively. The most productive countries were the United States (38%), followed by Germany (13%) and Japan (12%). Most articles were published in Br J Dermatol (13%) and J Invest Dermatol (11%). MeSH terms were evident in the prediction power of the number of article citations (correlation coefficient=0.45, t=4.99). CONCLUSIONS: The breakthrough was made by developing one dashboard to display PVBA100. MeSH terms can be used for predicting article citations in PVBA100. These visualizations of PVBA100 could be applied to future academic pursuits and applications in other academic disciplines.

Predicting the number of article citations on the topic of pemphigus vulgaris with the 100 top-cited articles since 2011: A protocol for systematic review and meta-analysis.

BACKGROUND: Pemphigus vulgaris (PV) is a rare autoimmune blistering disease characterized by intraepithelial and mucocutaneous blister formation and erosion. Numerous articles related to PV have been published. However, which articles have a tremendous influence is still unknown, and factors affecting article citation numbers remain unclear. We aimed to visualize the prominent entities using the top 100 most-cited articles on the topic of PV (T100PV), and investigate whether medical subject headings (i.e., MeSH terms) can be used to predict article citations. METHODS: By searching the PubMed Central (PMC) database, the T100PV abstracts since 2011 were downloaded. Citation analysis was performed to compare the dominant entities in article topics, authors, and research institutes using social network analysis (SNA) and Kano diagrams. We examined the MeSH prediction power against article citations using correlation coefficients (CCs). RESULTS: The most cited article (125 times) was authored by Ellebrecht from the University of Pennsylvania in the US. The most productive countries were Germany (28%) and the US (25%). Most articles were published in J Invest Dermatol (16%) and Br J Dermatol (10%). Kasperkiewicz (Germany) and the Normandie University (France) were the most cited authors and research institutes, respectively. The most frequently occurred MeSH terms were administration and dosage, immunology, and metabolism. MeSH terms were evident in the prediction power on the number of article citations (F = 19.77; P < .001). CONCLUSION: A breakthrough was achieved by developing dashboards to display the T100PV. MeSH terms can be used to predict the T100PV citations. These T100PV visualizations can be applied in future studies.

Suitability of h- and x-indices for evaluating authors' individual research achievements in a given short period of years: A bibliometric analysis.

BACKGROUND: The h-index of a researcher refers to the maximum number h of his/her publications that has at least h citations via the concept of the square area. The x-index is determined by the maximum area of a rectangle under the curve to interpret authors' individual research achievements (IRAs). However, the properties of both metrics have not been compared and discussed before. This study aimed to investigate whether both metrics of h- and x-index are suitable for evaluating IRAs in a short period of years. METHODS: By searching the PubMed database (Pubmed.com), we used the keyword "PLoS One" (journal) and downloaded 50,000 articles published in 2015 and 2016. A total of 146,346 citations were listed in PubMed Central and 27,035 authors(with h-index ≥1) were divided into 3 parts. Correlation coefficients among metrics (ie, AIF, h, g, Ag, and x-index) were examined. The bootstrapping method used for estimating 95% confidence intervals was applied to compare differences in metrics among author groups. The most cited authors and topic burst were visualized by social network analysis. The most prominent countries/areas were highlighted by the x-index and displayed via choropleth maps. RESULTS: Results demonstrated that, first, the h-index had the least relation to other metrics and failed to differentiate authors' IRAs among groups, particularly in a short time period. Second, the top 3 highest x-index for countries were the United States, China, and the UK but with the productivity-oriented feature. Third, the most cited medical subject headings (ie, MeSH terms) were genome, metabolome, and microbiology, and the most cited author was Lori Newman (whose x-index = 13.52, and h = 2) from Switzerland with the article (PMID = 26646541) cited 291 times. The need for the x-index combined with a visual map for displaying authors' IRAs was verified and recommended. CONCLUSIONS: We verified that the h-index failed to differentiate authors' IRAs among author groups in a short time period. The x-index combined with the Kano map is recommended in research for a better understanding of the authors' IRAs in other journals or disciplines, not just limited to the journal of PloS One as we did in this study.

Effects of Solanum undatum extract (SR-T100) on photocarcinogenesis and photoaging of actinic keratosis.

SR-T100 gel, containing solamargine extracted from Solanum undatum (synonym: Solanum incanum), had good therapeutic effects on actinic keratosis (AK) in human and ultraviolet B-induced papilloma in mice. This study aimed to investigate the immunohistochemical changes in the human skin after SR-T100 treatment. An immunohistochemical study was performed and the changes in photocarcinogenesis and photoaging markers after 16-week SR-T100 gel treatment were documented. SR-T100 gel treatment for 16 weeks resulted in complete remission in nine AK lesions and partial remission in four AK lesions. SR-T100 gel abolished the expression of mutant p53 and SOX2 and restored the expression of NOTCH1. Additionally, SR-T100 gel improved wrinkling in human skin, while restoring the expression of lamin B1 and increasing synthesis of new elastic fibers. SR-T100 gel had therapeutic effects on photocarcinogenesis and photoaging of photodamaged skin with AK.

Wwox Deficiency Causes Downregulation of Prosurvival ERK Signaling and Abnormal Homeostatic Responses in Mouse Skin.

Deficiency of tumor suppressor WW domain-containing oxidoreductase (WWOX) in humans and animals leads to growth retardation and premature death during postnatal developmental stages. Skin integrity is essential for organism survival due to its protection against dehydration and hypothermia. Our previous report demonstrated that human epidermal suprabasal cells express WWOX protein, and the expression is gradually increased toward the superficial differentiated cells prior to cornification. Here, we investigated whether abnormal skin development and homeostasis occur under Wwox deficiency that may correlate with early death. We determined that keratinocyte proliferation and differentiation were decreased, while apoptosis was increased in Wwox -/- mouse epidermis and primary keratinocyte cultures and WWOX-knockdown human HaCaT cells. Without WWOX, progenitor cells in hair follicle junctional zone underwent massive proliferation in early postnatal developmental stages and the stem/progenitor cell pools were depleted at postnatal day 21. These events lead to significantly decreased epidermal thickness, dehydration state, and delayed hair development in Wwox -/- mouse skin, which is associated with downregulation of prosurvival MEK/ERK signaling in Wwox -/- keratinocytes. Moreover, Wwox depletion results in substantial downregulation of dermal collagen contents in mice. Notably, Wwox -/- mice exhibit severe loss of subcutaneous adipose tissue and significant hypothermia. Collectively, our knockout mouse model supports the validity of WWOX in assisting epidermal and adipose homeostasis, and the involvement of prosurvival ERK pathway in the homeostatic responses regulated by WWOX.

Wwox deficiency leads to neurodevelopmental and degenerative neuropathies and glycogen synthase kinase 3ß-mediated epileptic seizure activity in mice.

Human WWOX gene resides in the chromosomal common fragile site FRA16D and encodes a tumor suppressor WW domain-containing oxidoreductase. Loss-of-function mutations in both alleles of WWOX gene lead to autosomal recessive abnormalities in pediatric patients from consanguineous families, including microcephaly, cerebellar ataxia with epilepsy, mental retardation, retinal degeneration, developmental delay and early death. Here, we report that targeted disruption of Wwox gene in mice causes neurodevelopmental disorders, encompassing abnormal neuronal differentiation and migration in the brain. Cerebral malformations, such as microcephaly and incomplete separation of the hemispheres by a partial interhemispheric fissure, neuronal disorganization and heterotopia, and defective cerebellar midline fusion are observed in Wwox-/- mice. Degenerative alterations including severe hypomyelination in the central nervous system, optic nerve atrophy, Purkinje cell loss and granular cell apoptosis in the cerebellum, and peripheral nerve demyelination due to Schwann cell apoptosis correspond to reduced amplitudes and a latency prolongation of transcranial motor evoked potentials, motor deficits and gait ataxia in Wwox-/- mice. Wwox gene ablation leads to the occurrence of spontaneous epilepsy and increased susceptibility to pilocarpine- and pentylenetetrazol (PTZ)-induced seizures in preweaning mice. We determined that a significantly increased activation of glycogen synthase kinase 3ß (GSK3ß) occurs in Wwox-/- mouse cerebral cortex, hippocampus and cerebellum. Inhibition of GSK3ß by lithium ion significantly abolishes the onset of PTZ-induced seizure in Wwox-/- mice. Together, our findings reveal that the neurodevelopmental and neurodegenerative deficits in Wwox knockout mice strikingly recapitulate the key features of human neuropathies, and that targeting GSK3ß with lithium ion ameliorates epilepsy.

Strategies by which WWOX-deficient metastatic cancer cells utilize to survive via dodging, compromising, and causing damage to WWOX-positive normal microenvironment.

Proapoptotic tumor suppressor WWOX is upregulated in the early stage of cancer initiation, which probably provides limitation to cancer growth and progression. Later, WWOX protein is reduced to enhance cancer cell growth, migration, invasiveness and metastasis. To understand how WWOX works in controlling cancer progression, here we demonstrate that apoptotic stress mediated by ectopic WWOX stimulated cancer cells to secrete basic fibroblast growth factor (bFGF) in order to support capillary microtubule formation. This event may occur in the cancer initiation stage. Later, when WWOX loss occurs in cancer cells, hyaluronidase production is then increased in the cancer cells to facilitate metastasis. We determined that inhibition of membrane hyaluronidase Tyr216-phosphorylated Hyal-2 by antibody suppresses cancer growth in vivo. WWOX-negative (WWOX-) cells dodged WWOX+cells in the microenvironment by migrating individually backward to avoid physical contacts and yet significantly upregulating the redox activity of WWOX+parental cells or other WWOX+cell types for causing apoptosis. Upon detecting the presence of WWOX+cells from a distance, WWOX- cells exhibit activation of MIF, Hyal-2, Eph, and Wnt pathways, which converges to MEK/ERK signaling and enables WWOX- cells to evade WWOX+cells. Inhibition of each pathway by antibody or specific chemicals enables WWOX- cells to merge with WWOX+cells. In addition, exogenous TGF-ß assists WWOX- cells to migrate collectively forward and merge with WWOX+cells. Metastatic WWOX- cancer cells frequently secrete high levels of TGF-ß, which conceivably assists them to merge with WWOX+cells in target organs and secure a new home base in the WWOX+microenvironment. Together, loss of WWOX allows cancer cells to develop strategies to dodge, compromise and even kill WWOX-positive cells in microenvironment.