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The transcription factor ΔNp63 plays a pivotal role in maintaining the integrity of stratified epithelial tissues by regulating the expression of distinct target genes involved in lineage specification, cell stemness, cell proliferation and differentiation. Here, we identified the ABC transporter subfamily member ABCC1 as a novel ΔNp63 target gene. We found that in immortalized human keratinocytes and in squamous cell carcinoma (SCC) cells, ∆Np63 induces the expression of ABCC1 by physically occupying a p63-binding site (p63 BS) located in the first intron of the ABCC1 gene locus. In cutaneous SCC and during the activation of the keratinocyte differentiation program, ∆Np63 and ABCC1 levels are positively correlated raising the possibility that ABCC1 might be involved in the regulation of the proliferative/differentiative capabilities of squamous tissue. However, we did not find any gross alteration in the structure and morphology of the epidermis in humanized hABCC1 knock-out mice. Conversely, we found that the genetic ablation of ABCC1 led to a marked reduction in inflammation-mediated proliferation of keratinocytes, suggesting that ABCC1 might be involved in the regulation of keratinocyte proliferation upon inflammatory/proliferative signals. In line with these observations, we found a significant increase in ABCC1 expression in squamous cell carcinomas (SCCs), a tumor type characterized by keratinocyte hyper-proliferation and a pro-inflammatory tumor microenvironment. Collectively, these data uncover ABCC1 as an additional ∆Np63 target gene potentially involved in those skin diseases characterized by dysregulation of proliferation/differentiation balance. [ABSTRACT FROM AUTHOR]

Giant cell tumor (GCT) is a benign, locally aggressive neoplasm with a high recurrence rate with common occurrence in the long bone and the cases in the maxillofacial region bone are very rare. Due to the paucity of the cases, there is not enough information available regarding the behavior of the tumor. Also, the differentiation of this aggressive lesion with the commonly occurring reactive giant cell lesions is crucial and needs more research. This study is pertaining to the review of literature of the cases of GCT in the oral cavity with their clinicopathological, radiographic, and biochemical analyses. Although there are no available studies regarding the immunohistochemical characteristics of this lesion, this study is the first step in this direction to differentiate this tumor better and identify the possible pathogenesis. [ABSTRACT FROM AUTHOR]

Abstract Background p63 is a transcription factor with intrinsic pioneer factor activity and pleiotropic functions. Transforming growth factor β (TGFβ) signaling via activation and cooperative action of canonical, SMAD, and non-canonical, MAP-kinase (MAPK) pathways, elicits both anti- and pro-tumorigenic properties, including cell stemness and invasiveness. TGFβ activates the ΔNp63 transcriptional program in cancer cells; however, the link between TGFβ and p63 in unmasking the epigenetic landscape during tumor progression allowing chromatin accessibility and gene transcription, is not yet reported. Methods Small molecule inhibitors, including protein kinase inhibitors and RNA-silencing, provided loss of function analyses. Sphere formation assays in cancer cells, chromatin immunoprecipitation and mRNA expression assays were utilized in order to gain mechanistic evidence. Mass spectrometry analysis coupled to co-immunoprecipitation assays revealed novel p63 interactors and their involvement in p63-dependent transcription. Results The sphere-forming capacity of breast cancer cells was enhanced upon TGFβ stimulation and significantly decreased upon ΔNp63 depletion. Activation of TGFβ signaling via p38 MAPK signaling induced ΔNp63 phosphorylation at Ser 66/68 resulting in stabilized ΔNp63 protein with enhanced DNA binding properties. TGFβ stimulation altered the ratio of H3K27ac and H3K27me3 histone modification marks, pointing towards higher H3K27ac and increased p300 acetyltransferase recruitment to chromatin. By silencing the expression of ΔNp63, the TGFβ effect on chromatin remodeling was abrogated. Inhibition of H3K27me3, revealed the important role of TGFβ as the upstream signal for guiding ΔNp63 to the TGFβ/SMAD gene loci, as well as the indispensable role of ΔNp63 in recruiting histone modifying enzymes, such as p300, to these genomic regions, regulating chromatin accessibility and gene transcription. Mechanistically, TGFβ through SMAD activation induced dissociation of ΔNp63 from NURD or NCOR/SMRT histone deacetylation complexes, while promoted the assembly of ΔNp63-p300 complexes, affecting the levels of histone acetylation and the outcome of ΔNp63-dependent transcription. Conclusions ΔNp63, phosphorylated and recruited by TGFβ to the TGFβ/SMAD/ΔNp63 gene loci, promotes chromatin accessibility and transcription of target genes related to stemness and cell invasion.

Introduction: Cervical cancer is one of the leading causes of cancer deaths. Liquid-based cervical cytology enables the detection and diagnosis of the disease at an early stage. p40 is a specific immunomarker that distinguishes squamous cell carcinomas from other cervical carcinomas with glandular and neuroendocrine differentiation. Aim: To determine the role of Liquid-based Cytology (LBC) in the early detection of premalignant and malignant lesions of the uterine cervix and its concordance with histopathological findings. Materials and Methods: This cross-sectional study was conducted at the Institute of Post Graduate Medical Education and Research (IPGME&R), Kolkata, West Bengal, India, over a period of one year and six months (from October 2020 to March 2022). Cervical samples were collected from a total of 200 females, and the materials were processed using the BD SurePath™ LBC method. The cytologically confirmed cases were biopsied, and histopathological concordance was established. Immunohistochemical (IHC) staining for p40 and p63 was performed using the peroxidase-antiperoxidase method to differentiate between premalignant, malignant squamous, and glandular lesions of the uterine cervix. For quantitative analysis, all cells were counted in 10 random fields at 400x magnification, and p63 and p40 were expressed as a percentage of positive cells per the total number of counted cells. Cases were considered positive if 5% or more of the tumour cell nuclei showed brown nuclear staining. The mean percentage positivity of p63 and p40 nuclear staining of all the cases in each category was calculated for non neoplastic and precursor/neoplastic lesions of the cervix. Results: Among the 200 cases evaluated by LBC, 169 (84.5%) were non neoplastic, and the remaining 31 (15.5%) were precursor/neoplastic lesions. The vast majority (126 cases, 63%) were inflammatory smears, followed by 20 cases (10%) that were Negative for Intraepithelial Lesion or Malignancy (NILM). Total of 40 cases were followed-up with biopsy and histopathology, and concordance with the cytological diagnosis was evaluated. Among 40 cases, 23 were precursor/neoplastic on histopathology, and seven cases that were non neoplastic on Histopathological Examination (HPE) had been correctly diagnosed on LBC, giving a concordance rate of 75%. The overall sensitivity, specificity, Positive Predictive Value (PPV), and Negative Predictive Value (NPV) of LBC were 92%, 46.6%, 74.1%, and 77.7%, respectively, considering histological diagnosis as the gold standard. Regarding IHC expression on histopathology, the mean positivity of p40 and p63 for non neoplastic lesions was 9.40% and 10.06%, and for precursor/neoplastic lesions, 47.44% and 46.4%, respectively. Adenocarcinoma-in-situ and adenocarcinoma were negative for both p40 and p63. Conclusion: Cervical cytology is a less invasive, cost-effective, and simple procedure to diagnose cervical epithelial cell abnormalities. Cytological-histopathological concordance revealed that LBC is a sensitive diagnostic method. The comparison between non neoplastic and neoplastic lesions of the cervix revealed a statistically significant difference with respect to the mean percent positivity of p40 and p63 IHC staining. Both of these markers can be used to differentiate squamous cell carcinoma from adenocarcinoma of the cervix.

p53 was discovered 45 years ago as an SV40 large T antigen binding protein, coded by the most frequently mutated TP53 gene in human cancers. As a transcription factor, p53 is tightly regulated by a rich network of post-translational modifications to execute its diverse functions in tumor suppression. Although early studies established p53-mediated cell-cycle arrest, apoptosis, and senescence as the classic barriers in cancer development, a growing number of new functions of p53 have been discovered and the scope of p53-mediated anti-tumor activity is largely expanded. Here, we review the complexity of different layers of p53 regulation, and the recent advance of the p53 pathway in metabolism, ferroptosis, immunity, and others that contribute to tumor suppression. We also discuss the challenge regarding how to activate p53 function specifically effective in inhibiting tumor growth without harming normal homeostasis for cancer therapy. p53 plays a central role in suppressing tumorigenesis. In this issue, Liu et al. review the complexity of different layers of p53 regulation, and the recent advance of the p53 pathway in metabolism, ferroptosis, immunity, and others that largely expand the scope of its anti-tumor activity. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are small RNA molecules that regulate more than 30% of genes in humans. Recent studies have revealed that miRNAs play a crucial role in tumorigenesis. Large sets of miRNAs in human tumors are under-expressed compared to normal tissues. Furthermore, experiments have shown that interference with miRNA processing enhances tumorigenesis. Multiple studies have documented the causal role of miRNAs in cancer, and miRNA-based anticancer therapies are currently being developed. This review primarily focuses on two key points: (1) miRNAs and their role in human cancer and (2) the regulation of tumor suppressors by miRNAs. The review discusses (a) the regulation of the tumor suppressor p53 by miRNA, (b) the critical role of the miR-144/451 cluster in regulating the Itch-p63-Ago2 pathway, and (c) the regulation of PTEN by miRNAs. Future research and the perspectives of miRNA in cancer are also discussed. Understanding these pathways will open avenues for therapeutic interventions targeting miRNA regulation. [ABSTRACT FROM AUTHOR]

Respiratory viruses cause airway inflammation, resulting in epithelial injury and repair. miRNAs, including miR-149-5p, regulate different pathological conditions. We aimed to determine how miR-149-5p functions in regulating pro-inflammatory IL-6 and p63, key regulators of airway epithelial wound repair, in response to viral proteins in bronchial (BEAS-2B) and alveolar (A549) epithelial cells. BEAS-2B or A549 cells were incubated with poly (I:C, 0.5 µg/mL) for 48 h or SARS-CoV-2 spike protein-1 or 2 subunit (S1 or S2, 1 μg/mL) for 24 h. miR-149-5p was suppressed in BEAS-2B challenged with poly (I:C), correlating with IL-6 and p63 upregulation. miR-149-5p was down-regulated in A549 stimulated with poly (I:C); IL-6 expression increased, but p63 protein levels were undetectable. miR-149-5p remained unchanged in cells exposed to S1 or S2, while S1 transfection increased IL-6 expression in BEAS-2B cells. Ectopic over-expression of miR-149-5p in BEAS-2B cells suppressed IL-6 and p63 mRNA levels and inhibited poly (I:C)-induced IL-6 and p63 mRNA expressions. miR-149-5p directly suppressed IL-6 mRNA in BEAS-2B cells. Hence, BEAS-2B cells respond differently to poly (I:C), S1 or S2 compared to A549 cells. Thus, miR-149-5p dysregulation may be involved in poly (I:C)-stimulated but not S1- or S2-stimulated increased IL-6 production and p63 expression in BEAS-2B cells. [ABSTRACT FROM AUTHOR]

Objective To explore the molecular mechanism of Gasdermin B (GSDMB) regulating the fate of intestinal epithelial cells. Methods The human GSDMB plasmid was overexpressed into two human intestinal epithelial cell lines (NCM460 and HT-29 cells) and human colon-derived organoids. Western blotting was used to confirm the efficiency of electroporation. Cell counting kit (CCK8), cell apoptosis, and cell cycle by flow cytometry were performed to analyze the effect of GSDMB overexpression on cell function. Transcriptome sequencing was used to analyze the downstream effector molecules of GSDMB. T test was used to compare the data between the two groups. Results The overexpression of GSDMB protein in the two intestinal epithelial cell lines was successfully reconstructed. The absorbance value (A) of human intestinal epithelial cells overexpressing GSDMB protein [NCM460 cells: (1.17±0.01), HT-29 cells: (0.96±0.06)] was significantly lower than that of blank control cells [NCM460 cells: (1.67±0.12), HT-29 cells: (1.24±0.07)] (t=7.24 and 5.46, P<0.05). The number of apoptotic cells in the GSDMB overexpression group [NCM460 cells: (12.03±1.55), HT-29 cells: (29.30±4.48)] was significantly higher than that in the blank group [NCM460 cells: (4.96±1.74), HT-29 cells: (6.95±3.42)](t=5.26 and 6.97, P<0.05). Cell cycle analysis showed that the ratio of cells at G0/G1 phase in the GSDMB overexpression group [NCM460 cells: (47.98±5.28)%, HT-29 cells: (38.04±3.45)%] was significantly lower than that in the control group [NCM460 cells: (59.54±3.90)%, HT-29 cells: (63.81±1.76) % (t=3.05 and 11.53, P<0.05). Transcriptome sequencing results showed that the dual specificity phosphatase 4 and 6 (DUSP4 and DUSP6) genes were significantly upregulated after GSDMB protein expression. Fluorescence quantitative PCR results confirmed that the relative expression levels of DUSP4 (2.45±0.15) and DUSP6 (4.34±0.22) in intestinal epithelial cells transfected with GSDMB were significantly higher than those in the control group (1.06±0.05 and 1.01±0.02)(t= 15.08 and 26.52, P<0.05). After GSDMB-expressing NCM460 cells were treated with the DUSP inhibitor BCI, the BCI treatment group had a significantly increased expression level of p-ERK compared to the control group [(1.14± 0.17) vs. (0.58±0.12)](t=5.42, P=0.002); the A value (1.84±0.07) and G0/G1 phase ratio (59.83±2.17)% in the BCI treatment group were significantly higher than those in the non-treatment group [(1.52±0.10) and (52.10± 2.23)%, and the number of apoptosis in the BCI treated group (7.60±0.56) was significantly lower than that in the untreated group (12.57±1.00) (t=4.71, 4.31, 7.52, P<0.05). TUNEL staining in human colon organoids showed a significant increase in apoptotic cells, and the relative expression level of DUSP6 protein (0.85±0.09) was significantly higher than that of the control group (0.21±0.04), accompanied by a decrease in p-ERK levels [(0.83± 0.18) vs. (0.19±0.06)], with statistical significance (t=11.95, P<0.001; t 6.56, P<0.001). Conclusion GSDMB may inhibit cell proliferation, induce cell cycle arrest, and promote apoptosis by upregulating dual specificity phosphatase DUSP6-mediated ERK phosphorylation, thus affecting the fate of intestinal epithelial cells. [ABSTRACT FROM AUTHOR]

Objective To investigate the expressions of H3.3G34W, p63 and SATB2 in giant cell tumor of bone (GCTB) and the effect and value of their combined application in the diagnosis of GCTB. Methods We collected the samples and medical records of 54 cases of GCTB and 83 cases of non-giant cell tumor of bone (14 cases of aneurysmal bone cyst, 16 cases of chondroblastoma and 53 cases of non-ossifying fibroma) diagnosed between 2020 and 2022 in the Department of Pathology of Honghui Hospital Affiliated to Xi'an Jiaotong University. The expressions of H3.3G34W, p63 and SATB2 were detected by EliVision immunohistochemical method. X² test was used to determine whether there are significant differences in the positive rates of H3.3G34W, p63 and SATB2 among all the groups. The combined diagnostic model including H3.3G34W, p63 and SATB2 was established by Logistic regression analysis, and the diagnostic value of the model was evaluated by ROC curve analysis. Results The positive rates of H3.3G34W, p63 and SATB2 in GCTB group were 81.5%, 90.7% and 92.6%, respectively; the positive rates in NGCTB group were 2.4%, 28.9% and 62.7%. Compared with NGCTB group, the age of GCTB group was significantly older [(41.222±14.849) vs. (16.566±9.439),P<0.001], and the prevalence was higher in women than in men (51.9% vs. 48.1%, P<0.001). In addition, compared with the NGCTB group, the positive rates of H3.3G34W (81.5% vs. 2.4%, P<0.001), p63 (90.7% vs. 28.9%, P<0.001) and SATB2 (92.6% vs. 62.7%, P<0.001) were significantly higher in the GCTB group. Univariate regression analysis built a univariate prediction model and ROC curve analysis showed that age (AUC=92.9%, P<0.001), sex (AUC=64.5%, P= 0.004), H3.3G34W positive rate (AUC=89.5%, P<0.001), p63 positive rate (AUC=80.9%, P<0.001) and SATB2 positive rate (AUC=65.0%, P=0.003) were independent predictors of diagnosis of giant cell tumor of bone. Multivariate regression analysis (Logistic) constructed a hybrid prediction model. ROC curve analysis suggested that the hybrid model showed better prediction value than the single factor model (AUC = 98.4%, P<0.001). Conclusion H3.3G34W, p63 and SATB2 are effective molecular markers for the diagnosis of GCTB, and their combined application can improve the prediction efficiency of the diagnosis of GCTB. [ABSTRACT FROM AUTHOR]

Introduction: Bladder cancer is a global disease, ranks as the fourth most prevalent cancer. The incidence and prevalence increase with age. Grade and aggressiveness have been found to be related with different genetic expression and mutation. Aims: To evaluate any relation of grade and invasiveness of urothelial cancer with varied expression of immune histochemical marker p63 and her2/neu. Materials and methods: The present study was a hospital based prospective cross-sectional study. This Study was conducted from July 2021 to April 2023 in the Urology department of a tertiary care hospital. Total 90 patients undergoing trans urethral resection of bladder tumour (TURBT) were included in this study. Result: It was found that, patients who had decreased p63 expression had high grade in tumours (93.1%) compared to patients who were expressing normal p63 (32.8%) and this was statistically significant (p < 0.0001). Tumours with decreased p63 also appeared to be more invasive, 62.1% were found to be muscle invasive. Tumours with her2 neu expression found to be more aggressive in nature, 85.7% had high grade features and 53.6% were muscle invasive. Conclusion: Our findings suggest that immunohistochemical expression of HER2/neu positive and decreased p63 expression were associated with high grade and invasiveness in case of bladder carcinoma. [ABSTRACT FROM AUTHOR]

Tumour protein 63 (p63) is a transcription factor of the p53 gene family, encoded by the TP63 gene located at chromosome 3q28, which regulates the activity of genes involved in growth and development of the ectoderm and derived tissues. p63 protein is normally expressed in the nuclei of the basal cell layer of glandular organs, including breast, in squamous epithelium and in urothelium. p63 immunohistochemical (IHC) staining has several applications in diagnostic breast pathology. It is commonly used to demonstrate myoepithelial cells at the epithelial stromal interface to differentiate benign and in situ lesions from invasive carcinoma and to characterize and classify papillary lesions including the distinction of breast intraduct papilloma from skin hidradenoma. p63 IHC is also used to identify and profile lesions showing myoepithelial cell and/or squamous differentiation, e.g. adenomyoepithelioma, salivary gland‐like tumours including adenoid cystic carcinoma, and metaplastic breast carcinoma including low‐grade adenosquamous carcinoma. This article reviews the applications of p63 IHC in diagnostic breast pathology and outlines a practical approach to the diagnosis and characterization of breast lesions through the identification of normal and abnormal p63 protein expression. The biology of p63, the range of available antibodies with emphasis on staining specificity and sensitivity, and pitfalls in interpretation are also discussed. The TP63 gene in humans, which shows a specific genomic structure, resulting in either TAp63 (p63) isoform or ΔNp63 (p40) isoform. As illustrated in the figure, both isoforms contain a DNA‐binding domain (Orange box) and an oligomerization domain (Grey box). TAp63 contains an N‐terminal transactivation (TA) domain (Green box), while ΔNp63 has an alternative terminus (Yellow box). Antibodies against conventional pan‐p63 (TP63) bind to the DNA binding domain common to both isoforms (TAp63 and p40) and does not distinguish between them. Antibodies against TAp63 bind to the N‐terminal TA domain, while antibodies specific to ΔNp63 (p40) bind to the alternative terminus. Each isoform has variant isotypes (α, β, γ, δ, and ε). [ABSTRACT FROM AUTHOR]

Background: It is a well-recognized fact that abnormal cell proliferation plays a crucial role in the development of odontogenic lesions. p53 is a tumour-suppressor gene which assists in cell cycle regulation and p63 is a homolog of p53 responsible for ectodermal differentiation and maintenance of stratified epithelial progenitor-cell. Analysing the tissue expression of p53 and p63 in odontogenic lesions may provide us with an insight into their potential role in the development of these lesions. Objective: The objective is to study the expression of p53 and p63 in selected odontogenic lesions using immunohistochemistry. Materials and Methods: Formalin-fixed paraffin-embedded tissues of 15 ameloblastomas, 10 adenomatoid odontogenic tumours (AOT), 15 odontogenic keratocysts (OKCs), 10 dentigerous cysts (DCs) along with 10 cases of normal mucosa were retrieved from the departmental archives. These specimens were then subjected to immunohistochemical staining using p53 and p63 oncoproteins. Results: p53 and p63 immune-expression showed mainly intranuclear localization. The mean positivity of p53 in ameloblastoma (59.45%) and OKC (26.38%) was significantly higher than AOT (6.77%) and DC (4%). In contrast, there was no significant difference in the positivity of p63 in between ameloblastoma (77.55%), AOT (69.50%), OKC (76.47%), and DC (50.69%). Conclusion: p53 expression can be correlated with the clinical behaviour of the odontogenic lesions and it can be used as a prognostic marker in odontogenic cysts and tumours. In contrast, p63 expression does not corelate with the biological behaviour of odontogenic lesions. [ABSTRACT FROM AUTHOR]

Introduction: Breast carcinoma is leading cause of cancer death in women. Breast lesions constitute heterogeneous group of diseases with wide variety of etiologies ranging from inflammatory-benign- malignant lesions. There are several reported markers for immunohistochemical detection of myoepithelial cells. Smooth muscle specific proteins, such as smooth muscle actin, smooth muscle myosin heavy chain, calponin and h-caldesmon are used to highlight myoepithelium. p63 antibody is myoepithelial cell marker that selectively stains nuclei. It is negative in stromal, myofibroblastic and adipocytic cells. Aims and Objectives: The aim of this study was to establish role of p63 expression in distinguishing benign breast lesions, premalignant lesions and malignant tumors of breast. Materials and Methods: 30 cases were selected from core biopsy, lumpectomy and mastectomy specimens of breast received at department of Pathology, Chalmeda Anand Rao Institute of Medical Sciences during the period of January 2021 to December 2021 and were studied prospectively. All specimens were processed according to CAP protocol and reported . Immunohistochemistry was performed to determine p63 expression in those specimens. p63expression was evaluated as continuous positive/discontinuous positive/negative. Ethical Approval: This study was reviewed and approved by institute ethics committee, CAIMS, Karimnagar. Results: Among total 30 cases, 18 cases (60%) were benign lesions and all were positive for p63 expression. 3 cases (9.99%) were premalignant and were least positive for p63 expression. All malignant cases 9 cases (29.99%) were negative for p63 expression. Conclusion: The Positive correlation was seen between histomorphological features and p63 scoring in all the lesions, So p63 is good Immunohistochemical marker for evaluating breast lesions. [ABSTRACT FROM AUTHOR]

Objective: p63 is a transcription factor involved in multiple biological functions. In the liver, the TAp63 isoform induces lipid accumulation in hepatocytes. However, the role of liver TAp63 in the progression of metabolic dysfunction-associated steatohepatitis (MASH) with fibrosis is unknown. Methods: We evaluated the hepatic p63 levels in different mouse models of steatohepatitis with fibrosis induced by diet. Next, we used virogenetic approaches to manipulate the expression of TAp63 in adult mice under diet-induced steatohepatitis with fibrosis and characterized the disease condition. Finally, we performed proteomics analysis in mice with overexpression and knockdown of hepatic TAp63. Results: Levels of TAp63, but not of ΔN isoform, are increased in the liver of mice with diet-induced steatohepatitis with fibrosis. Both preventive and interventional strategies for the knockdown of hepatic TAp63 significantly ameliorated diet-induced steatohepatitis with fibrosis in mice fed a methionine- and choline-deficient diet (MCDD) and choline deficient and high fat diet (CDHFD). The overexpression of hepatic TAp63 in mice aggravated the liver condition in mice fed a CDHFD. Proteomic analysis in the liver of these mice revealed alteration in multiple proteins and pathways, such as oxidative phosphorylation, antioxidant activity, peroxisome function and LDL clearance. Conclusions: These results indicate that liver TAp63 plays a critical role in the progression of diet-induced steatohepatitis with fibrosis, and its inhibition ameliorates the disease.

PurposeThis review will discuss the role of vitamin D and calcium signaling in the epidermal wound response with particular focus on the stem cells of the epidermis and hair follicle that contribute to the wounding response.MethodsSelected publications relevant to the mechanisms of wound healing in general and the roles of calcium and vitamin D in wound healing in particular were reviewed.ResultsFollowing wounding the stem cells of the hair follicle and interfollicular epidermis are activated to proliferate and migrate to the wound where they take on an epidermal fate to re-epithelialize the wound and regenerate the epidermis. The vitamin D and calcium sensing receptors (VDR and CaSR, respectively) are expressed in the stem cells of the hair follicle and epidermis where they play a critical role in enabling the stem cells to respond to wounding. Deletion of Vdr and/or Casr from these cells delays wound healing. The VDR is regulated by co-regulators such as the Med 1 complex and other transcription factors such as Ctnnb (beta-catenin) and p63. The formation of the Cdh1/Ctnn (E-cadherin/catenin) complex jointly stimulated by vitamin D and calcium plays a critical role in the activation, migration, and re-epithelialization processes.ConclusionVitamin D and calcium signaling are critical for the ability of epidermal and hair follicle stem cells to respond to wounding. Vitamin D deficiency with the accompanying decrease in calcium signaling can result in delayed and/or chronic wounds, a major cause of morbidity, loss of productivity, and medical expense.

There is an ongoing need for biomarkers that could reliably predict the outcome of BC and that could guide the management of this disease. In this setting, we aimed to explore the prognostic value of the transcription factor P63 in patients with muscle-invasive bladder cancer (MIBC) having undergone radical cystectomy. The correlation between P63 expression and clinicopathological features (tumor stage, nodes involvement, patterns of muscularis propria invasion, papillary architecture, anaplasia, concomitant carcinoma in situ, lymphovascular invasion, perineural invasion, necrosis) and molecular subtyping (basal and luminal type tumors) was tested in 65 radical cystectomy specimens and matched with cancer-specific survival (CSS) and overall survival (OS). P63-negative tumors displayed significantly higher rates of pattern 2 of muscularis propria invasion (50% vs. 14%, p = 0.002) and variant histology (45% vs. 19%, p = 0.022) compared to P63-positive ones. According to the combined expression of CK5/6 and CK20 (Algorithm #1), P63-positive and P63-negative tumors were mostly basal-like and double-negative, respectively (p = 0.004). Using Algorithm #2, based on the combined expression of CK5/6 and GATA3, the vast majority of tumors were luminal overall and in each group (p = 0.003). There was no significant difference in CSS and OS between P63-positive and P63-negative tumors, but the former featured a trend towards longer OS. Though associated with pathological features harboring negative prognostic potential, P63 status as such failed to predict CSS and OS. That said, it may contribute to better molecular subtyping of MIBC.

Human papillomavirus type 8 (HPV8), a cutaneous genus beta HPV type, has co-carcinogenic potential at sun-exposed sites in patients suffering from the inherited skin disease epidermodysplasia verruciformis (EV). We had previously shown that Langerhans cells responsible for epithelial immunosurveillance were strongly reduced at infected sites and that the HPV8 E7 protein interferes with the CCAAT/enhancer-binding protein (C/EBP)β to suppress the Langerhans cell chemokine CCL20. At the same time, however, we observed that EV lesions are heavily infiltrated with inflammatory immune cells, which is similar to the situation in HPV8 E6 transgenic mice. To identify critical inflammatory factors, we used a broad multiplex approach and found that the monocyte attracting chemokine CCL2 was significantly and strongly induced by HPV8 E6 but not E7-expressing HaCaT cells, which were used as a model for UV-damaged skin keratinocytes. Conditioned media from HPV8 E6-expressing keratinocytes enhanced CCL2-receptor (CCR2)-dependent monocyte recruitment in vitro, and macrophages predominated in the stroma but were also detected in the epidermal compartment of EV lesions in vivo. CCL2 induction by HPV8 E6 was even stronger than stimulation with the proinflammatory cytokine TNF-α, and both HPV8 E6 and TNF-α resulted in substantial suppression of the transcription factor C/EBPα. Using RNAi-mediated knockdown and overexpression approaches, we demonstrated a mechanistic role of the recently identified C/EBPα/miR-203/p63 pathway for HPV8 E6-mediated CCL2 induction at protein and transcriptional levels. Epithelial co-expression of p63 and CCL2 was confirmed in HPV8 E6-expressing organotypic air-liquid interface cultures and in lesional EV epidermis in vivo. In summary, our data demonstrate that HPV8 oncoproteins actively deregulate epidermal immune homeostasis through modulation of C/EBP factor-dependent pathways. While HPV8 E7 suppresses immunosurveillance required for viral persistence, the present study provides evidence that E6 involves the stemness-promoting factor p63 to support an inflammatory microenvironment that may fuel carcinogenesis in EV lesions. [ABSTRACT FROM AUTHOR]

Histological staining is the primary method for confirming cancer diagnoses, but certain types, such as p63 staining, can be expensive and potentially damaging to tissues. In our research, we innovate by generating p63-stained images from H&E-stained slides for metaplastic breast cancer. This is a crucial development, considering the high costs and tissue risks associated with direct p63 staining. Our approach employs an advanced CycleGAN architecture, xAI-CycleGAN, enhanced with context-based loss to maintain structural integrity. The inclusion of convolutional attention in our model distinguishes between structural and color details more effectively, thus significantly enhancing the visual quality of the results. This approach shows a marked improvement over the base xAI-CycleGAN and standard CycleGAN models, offering the benefits of a more compact network and faster training even with the inclusion of attention. [ABSTRACT FROM AUTHOR]

The alteration of progenitor/stem cells present in the airway epithelium has been observed in patients with COPD. Smoking exposure induces remodeling patterns in bronchial progenitor cells (BPCs), encompassing squamous metaplasia, hyperplasia of basal and of mucus-secreting cells, and the depletion of ciliated and non-mucous secretory cells. Our aim was to assess the expression of p63 and vimentin as potential markers of airway remodeling and the regulation of stem cell populations in obstructive and neoplastic lung disease patients. A retrospective single-center observational study was conducted, including patients undergoing bronchoscopy with bronchial biopsies for suspected lung cancer. p63 and vimentin expression were evaluated via immunohistochemical analysis. There were 25 patients, of which 21 with COPD were included, and 17 were diagnosed with lung cancer. We observed that FEV1% was negatively correlated with p63+ basal cell number (r = −0.614, p = 0.019) and positively correlated with vimentin expression (r = 0.670; p = 0.008). p63 was significantly higher in biopsies from the trachea and main bronchi compared to more distal areas (p = 0.040), whereas vimentin was prevalent in the more distal areas (p = 0.042). Our preliminary data suggest the initial evidence of structural changes in BPCs among patients with COPD and lung cancer. Further research efforts are warranted to investigate additional morphologic and functional respiratory parameters in these patients. [ABSTRACT FROM AUTHOR]

Objectives: Meningiomas, a common neoplasm of the central nervous system, is a widely studied meningeal tumor. According to the World Health Organization (WHO) 2021 classification of meningiomas, there are 15 subtypes that have been grouped into grades 1, 2, and 3. The WHO grade 1 meningiomas are generally grouped as benign while the WHO grades 2 and 3 tumors are grouped as malignant. Progesterone receptors and P63 are common immunohistochemical markers that have proven useful in the diagnosis, grading, and prognostication of many neoplasms such as breast carcinoma, prostate carcinoma, and gastrointestinal tumors in histopathology practice. The application of these immunohistochemical markers to the grading of meningiomas has been reported and their usefulness documented in reports from Africa, Europe, North America, South America, and Asia. This study, therefore, seeks to determine if these findings are applicable to the meningiomas seen in an African population. Materials and Methods: A 10-year review of results and histologically diagnosed cases of meningiomas received in the Department of Morbid Anatomy, University of Nigeria, Enugu. Immunostaining for progesterone receptors (PgRs) and P63 were done and results compared with histologic grades. Results: The three WHO grades of meningioma were assessed in this study. M: F ratio was 1:1.4 and peak age was 41-50 years age range (SD ± 16.54). The majority of the cases were WHO grade 1 (86.1%) while the WHO grades 2 and 3 tumors were 8% and 5.9%, respectively. The fibrous variant was the most common subtype (27.1%). There was no correlation between progesterone receptor and P63 immunopositivity to the WHO grades of meningioma (P = 0.112 and P = 0.138, respectively). Conclusion: Our study showed that progesterone receptors and P63 immunopositivity did not correlate with the WHO grades of meningiomas. This may be due to the predominant variant of meningioma seen in this study. These findings indicate that PgR antagonist may not be an effective alternative for treatment in patients with inoperable meningiomas. Furthermore, P63 immunopositivity may not be a sufficient grading tool for managing meningiomas in our population. [ABSTRACT FROM AUTHOR]

Merkel cell carcinoma (MCC) is a cutaneous neuroendocrine tumor, and more than 90% of feline MCC cases test positive for Felis catus papillomavirus type 2 (FcaPV2). In the present study, basal cell markers p40, p63, and p73 and the stem cell marker SOX2 and cytokeratin 14 (CK14) were immunohistochemically examined in normal fetal, infant, and adult feline skin tissues. The expression of these proteins was examined in tumors positive for FcaPV2, including MCC, basal cell carcinoma (BCC), Bowenoid in situ carcinoma (BISC), and squamous cell carcinoma (SCC). Infant and adult feline skin tissues had mature Merkel cells, which were CK14-, CK18+, CK20+, SOX2+, synaptophysin+ and CD56+, while fetal skin tissue had no mature Merkel cells. MCC was immunopositive for p73, CK18, and SOX2 in 32/32 cases, and immunonegative for CK14 in 31/32 cases and for p40 and p63 in 32/32 cases. These results indicate that MCC exhibits different immunophenotypes from Merkel cells (p73-) and basal cells (p40+, p63+, and SOX2-). In contrast, all 3 BCCs, 1 BISC, and 2 SCCs were immunopositive for the basal cell markers p40, p63, and p73. The life cycle of papillomavirus is closely associated with the differentiation of infected basal cells, which requires the transcription factor p63. Changes in p63 expression in FcaPV2-positive MCC may be associated with unique cytokeratin expression patterns (CK14-, CK18+, and CK20+). Furthermore, SOX2 appears to be involved in Merkel cell differentiation in cats, similar to humans and mice. [ABSTRACT FROM AUTHOR]

Abstract Background Limbal stem cells (LSCs) are crucial for the regeneration of the corneal epithelium in patients with limbal stem cell deficiency (LSCD). Thus, LSCs during cultivation in vitro should be in highly homogeneous amounts, while potency and expression of stemness without tumorigenesis would be desirable. Therefore, further characterization and safety evaluation of engineered limbal grafts is required to provide safe and high-quality therapeutic applications. Methods After in vitro expansion, LSCs undergo laboratory characterization in a single-cell suspension, cell culture, and in limbal grafts before transplantation. Using a clinically applicable protocol, the data collected on LSCs at passage 1 were summarized, including: identity (cell size, morphology); potency (yield, viability, population doubling time, colony-forming efficiency); expression of putative stem cell markers through flow cytometry, immunofluorescence, and immunohistochemistry. Then, mitotic chromosome stability and normal mitotic outcomes were explored by using live-cell imaging. Finally, impurities, bacterial endotoxins and sterility were determined. Results Expression of the stemness marker p63 in single-cell suspension and in cell culture showed high values by different methods. Limbal grafts showed p63-positive cells (78.7 ± 9.4%), Ki67 proliferation (41.7 ± 15.9%), while CK3 was negative. Impurity with 3T3 feeder cells and endotoxins was minimized. We presented mitotic spindles with a length of 11.40 ± 0.54 m and a spindle width of 8.05 ± 0.55 m as new characterization in LSC culture. Additionally, live-cell imaging of LSCs (n = 873) was performed, and only a small fraction

Abstract Background Head and neck squamous cell carcinoma (HNSCC) is characterized by high proliferation and limited differentiation. The altered expression of the p53 family members, and specifically of p63, represents a pivotal event in the pathogenesis of HNSCC. Physiologically, p63 affects metabolism through the direct transactivation of the enzyme hexokinase 2, and subsequently controls the proliferation of epithelial cells; nonetheless, its role in cancer metabolism is still largely unclear. The high energetic demand of cancer and the consequent needs of a metabolic reshape, also involve the serine and glycine catabolic and anabolic pathways, including the one carbon metabolism (OCM), to produce energetic compounds (purines) and to maintain cellular homeostasis (glutathione and S-adenosylmethionine). Results The involvement in serine/glycine starvation by other p53 family members has been reported, including HNSCC. Here, we show that in HNSCC p63 controls the expression of the enzymes regulating the serine biosynthesis and one carbon metabolism. p63 binds the promoter region of genes involved in the serine biosynthesis as well as in the one carbon metabolism. p63 silencing in a HNSCC cell line affects the mRNA and protein levels of these selected enzymes. Moreover, the higher expression of TP63 and its target enzymes, negatively impacts on the overall survival of HNSCC patients. Conclusion These data indicate a direct role of p63 in the metabolic regulation of HNSCC with significant clinical effects.

The transcription factor ΔNp63 plays a pivotal role in maintaining the integrity of stratified epithelial tissues by regulating the expression of distinct target genes involved in lineage specification, cell stemness, cell proliferation and differentiation. Here, we identified the ABC transporter subfamily member ABCC1 as a novel ΔNp63 target gene. We found that in immortalized human keratinocytes and in squamous cell carcinoma (SCC) cells, ∆Np63 induces the expression of ABCC1 by physically occupying a p63-binding site (p63 BS) located in the first intron of the ABCC1 gene locus. In cutaneous SCC and during the activation of the keratinocyte differentiation program, ∆Np63 and ABCC1 levels are positively correlated raising the possibility that ABCC1 might be involved in the regulation of the proliferative/differentiative capabilities of squamous tissue. However, we did not find any gross alteration in the structure and morphology of the epidermis in humanized hABCC1 knock-out mice. Conversely, we found that the genetic ablation of ABCC1 led to a marked reduction in inflammation-mediated proliferation of keratinocytes, suggesting that ABCC1 might be involved in the regulation of keratinocyte proliferation upon inflammatory/proliferative signals. In line with these observations, we found a significant increase in ABCC1 expression in squamous cell carcinomas (SCCs), a tumor type characterized by keratinocyte hyper-proliferation and a pro-inflammatory tumor microenvironment. Collectively, these data uncover ABCC1 as an additional ∆Np63 target gene potentially involved in those skin diseases characterized by dysregulation of proliferation/differentiation balance.

Background: Limbal stem cells (LSCs) are crucial for the regeneration of the corneal epithelium in patients with limbal stem cell deficiency (LSCD). Thus, LSCs during cultivation in vitro should be in highly homogeneous amounts, while potency and expression of stemness without tumorigenesis would be desirable. Therefore, further characterization and safety evaluation of engineered limbal grafts is required to provide safe and high-quality therapeutic applications. Methods: After in vitro expansion, LSCs undergo laboratory characterization in a single-cell suspension, cell culture, and in limbal grafts before transplantation. Using a clinically applicable protocol, the data collected on LSCs at passage 1 were summarized, including: identity (cell size, morphology); potency (yield, viability, population doubling time, colony-forming efficiency); expression of putative stem cell markers through flow cytometry, immunofluorescence, and immunohistochemistry. Then, mitotic chromosome stability and normal mitotic outcomes were explored by using live-cell imaging. Finally, impurities, bacterial endotoxins and sterility were determined. Results: Expression of the stemness marker p63 in single-cell suspension and in cell culture showed high values by different methods. Limbal grafts showed p63-positive cells (78.7 ± 9.4%), Ki67 proliferation (41.7 ± 15.9%), while CK3 was negative. Impurity with 3T3 feeder cells and endotoxins was minimized. We presented mitotic spindles with a length of 11.40 ± 0.54 m and a spindle width of 8.05 ± 0.55 m as new characterization in LSC culture. Additionally, live-cell imaging of LSCs (n = 873) was performed, and only a small fraction < 2.5% of aberrant interphase cells was observed; 2.12 ± 2.10% of mitotic spindles exhibited a multipolar phenotype during metaphase, and 3.84 ± 3.77% of anaphase cells had a DNA signal present within the spindle midzone, indicating a chromosome bridge or lagging chromosome phenotype. Conclusion: This manuscript provides, for the first time, detailed characterization of the parameters of fidelity of the mitotic process and mitotic spindle morphologies of LSCs used in a direct clinical application. Our data show that p63-positive CK3-negative LSCs grown in vitro for clinical purposes undergo mitotic processes with extremely high fidelity, suggesting high karyotype stability. This finding confirms LSCs as a high-quality and safe therapy for eye regeneration in humans. [ABSTRACT FROM AUTHOR]

Cultured epidermal cell sheets and bioengineered skin equivalents are widely used for skin reconstruction. Cryostorage of keratinocytes is one of the steps of the bioengineered products manufacture; therefore, it is important to study its effect on the viability of these cells. We investigated the preservation of morphogenetic potential by keratinocytes after cryostorage and found that cryostorage did not alter the dynamics of their proliferation. The population of keratinocytes is known to be highly heterogeneous in terms of proliferative and regenerative abilities. Our analysis of the pattern of keratinocyte death in suspension before and after cryostorage revealed the most vulnerable population with a high level of α6 integrin expression and a low level of transferrin receptor expression. This data must be taken into account in the course of tissue engineering. [ABSTRACT FROM AUTHOR]

Background/Aims: The Ectrodactyly-Ectodermal dysplasia-Clefting (EEC) and Ankyloblepharon–ectodermal defect–cleft lip/palate (AEC) syndromes are rare autosomal dominant diseases caused by heterozygous mutations in the p63 gene. Patients are characterized by abnormalities of the skin, teeth, and hair and have limb defects, orofacial clefting and ectodermal dysplasia. In addition, they often show ocular surface alterations, leading to progressive corneal clouding and eventually blindness. Here, we present 8 cases describing patients affected by EEC (n = 6, with 5 sporadic and 1 familial cases) and AEC (n = 2, both sporadic cases) syndromes. We attempt to provide a description of the ocular disease progression over the years. Methods: Clinical examinations and monitoring of ocular parameters for the assessment of limbal stem cell deficiency were constantly performed on patients between 2009 and 2023. Quantitative data and comparison with existing cases described in the literature are reported. Results: The therapies supplied to patients were essential for the management of the symptoms, but unfortunately did not halt the progression of the pathology. Conclusions: A constant monitoring of the patients would help avoid the sudden worsening of symptoms. If the progression of the disease slows down, it would allow for the development of newer therapeutic strategies aimed at correcting the genetic defect. [ABSTRACT FROM AUTHOR]

Background: Extramammary Paget disease (EMPD), pagetoid squamous cell carcinoma in situ (PSCCIS), and Paget disease of the breast (PD) are intraepidermal carcinomas with overlapping histopathologic features. CK7 and CAM5.2 stains are frequently utilized to distinguish PSCCIS from EMPD and PD. However, some cases of PSCCIS can stain positively for CAM5.2 and CK7, indicating a potential pitfall with these stains. p63 has been shown to distinguish PSCCIS from EMPD. We assessed p63 staining in PD and compared it to p63 staining of PSCCIS and EMPD. Methods: A retrospective search for 15 examples each of PSCCIS, EMPD, and PD with remaining tissue in the paraffin block was performed. The diagnosis was confirmed by a board‐certified dermatopathologist and immunostaining for p63, CK7, and CAM5.2 was performed. Staining >55% was scored as positive. Staining <55% was scored as negative and an approximate percentage of positive cells was recorded. Results: Diffuse nuclear expression for p63 was detected in 100% (15/15) of PSCCIS cases, 0% (0/15) of PD cases, and 0% (0/15) of EMPD cases. CK7 and CAM5.2 stains were positive in 100% of PD. CAM5.2 was positive in 100% of EMPD and CK7 was positive in 93% of EMPD. CAM5.2 was positive in 0% of PSCCIS biopsy specimens, but partial staining was seen in 20%. CK7 was positive in 13%, but partial staining was seen in 47%. Conclusions: p63 immunostaining is a highly sensitive and specific method for differentiating between PSCCIS and PD or EMPD. While CAM5.2 and CK7 are also useful ancillary stains in this differential diagnosis, false‐positive and false‐negative staining occurs with these two markers. [ABSTRACT FROM AUTHOR]

Background: Head and neck squamous cell carcinoma (HNSCC) is characterized by high proliferation and limited differentiation. The altered expression of the p53 family members, and specifically of p63, represents a pivotal event in the pathogenesis of HNSCC. Physiologically, p63 affects metabolism through the direct transactivation of the enzyme hexokinase 2, and subsequently controls the proliferation of epithelial cells; nonetheless, its role in cancer metabolism is still largely unclear. The high energetic demand of cancer and the consequent needs of a metabolic reshape, also involve the serine and glycine catabolic and anabolic pathways, including the one carbon metabolism (OCM), to produce energetic compounds (purines) and to maintain cellular homeostasis (glutathione and S-adenosylmethionine). Results: The involvement in serine/glycine starvation by other p53 family members has been reported, including HNSCC. Here, we show that in HNSCC p63 controls the expression of the enzymes regulating the serine biosynthesis and one carbon metabolism. p63 binds the promoter region of genes involved in the serine biosynthesis as well as in the one carbon metabolism. p63 silencing in a HNSCC cell line affects the mRNA and protein levels of these selected enzymes. Moreover, the higher expression of TP63 and its target enzymes, negatively impacts on the overall survival of HNSCC patients. Conclusion: These data indicate a direct role of p63 in the metabolic regulation of HNSCC with significant clinical effects. [ABSTRACT FROM AUTHOR]

The NONO::TFE3 fusion has been described in MiT family translocation renal cell carcinomas as well as extracutaneous perivascular epithelioid cell tumors (PEComas). PEComas are known to express myogenic and melanocytic markers but SOX10 and p63 positivity has never been reported. We report two primary cutaneous tumors that morphologically and molecularly fit PEComas, both harboring the NONO::TFE3 fusion, but with an unusual immunophenotype of SOX10 and p63 positivity. One case was on an 80‐year‐old male's finger, and the other one was on a 72‐year‐old female's thigh. Both were well‐circumscribed multinodular dermal tumors composed of nests of monotonous epithelioid to spindled cells with pale to vacuolated cytoplasm, some of which were arranged around blood vessels. Both tumors were positive for SOX10, S100, and p63, focally positive for Melan‐A, and negative for myogenic markers. There are very little data regarding the molecular findings of primary cutaneous PEComas. While the NONO::TFE3 fusion has been identified in extracutaneous PEComas, it has never been reported in primary cutaneous cases. We believe these cases represent a previously undescribed subtype of cutaneous tumor which shows some immunophenotypic expression of melanocytic markers and we named these cases NONO::TFE3 fusion cutaneous epithelioid and spindle cell tumor. Further, we raise the question of whether this tumor should fall under the rubric of PEComa because of its morphology, partial expression of melanocytic markers, and the presence of the NONO::TFE3 fusion, or whether these tumors represent a separate novel class of tumors since the immunophenotypic expression of SOX10 and p63 is unusual for PEComas. [ABSTRACT FROM AUTHOR]

Background: Two subtypes of vulvar squamous cell carcinomas (VSCC) have previously been defined. The more common keratinising type typically occurs in older women (50- 70 years), is generally associated with lichen sclerosis and/or differentiated vulvar intraepithelial neoplasia (dVIN), and is often associated with p53 tumor suppressor gene mutations. The other subtype is more common in younger women and primarily associated with human papilloma virus (HPV) infection, and a common precursor is usual-type vulvar intraepithelial neoplasia (uVIN) of the basaloid or warty type. To investigate the prognostic significance of HPV status in vulvar squamous cell carcinomas (VSCC) and to determine whether preoperative determination of p16 or p53 status would have clinical relevance. Materials and methods: This is prospective and descriptive study conducted in the Department of Pathology, Government Medical College, Siddipet over a period of 1 year. Immunohistochemistry was performed by using 5 μm paraffin sections which were deparaffinized in xylene (three times for five minutes) and rehydrated in decreasing concentrations of ethanol (100%, 96%; two times each for ten minutes) followed by washing in deionised H2O for one minute. To unmask the p16 antigen the slides were covered with 0.01 M sodium citrate buffer (pH 6.0) and placed on a hot plate (950C) for 10 minutes. After cooling down the specimens were rinsed briefly in deionised H2O (three times). The specific primary (mouse monoclonal) antibody (p16, clone E6H4) was applied in a dilution of 1:50 overnight at 40C. Result: HPV types and status in correlation with clinical parameters and expression of p16. 70 out of 70 patients with PCV could be evaluated for HPV status. 25 were positive for high-risk HPV and 45 were HPV negative. The majority (16 out of 26, 64%) of HPVpositive patients were positive for HPV16. The others were positive for HPV45 (4 patients, 16%), HPV18 (2 patient, 8%), HPV35 (1 patient), HPV56 (1 patient), and HPV68 (1 patient). Human papillomavirus positivity was significantly correlated with strong p16 expression (p= 0.045). In all, 7 out of the 59 HPV-negative patients were negative for p16 immunostaining, while the remaining 83% showed varying expression: 31 out of 45 (68.9%) showed moderate or strong p16 expression. Conclusion: The vast majority of HPV positive vaginal cancers showed p16 overexpression, suggesting active involvement of HPV in the malignant transformation process. HPV vaccines will help prevent some of the primary female genital cancers associated with HPV type 16. More indepth studies are needed to understand the molecular carcinogenesis pathway in these p16-negative tumors and to improve outcomes for this population. [ABSTRACT FROM AUTHOR]

Self-renewing, damage–repair and differentiation of mammalian stratified squamous epithelia are subject to tissue homeostasis, but the regulation mechanisms remain elusive. Here, we investigate the esophageal squamous epithelial tissue homeostasis in vitro and in vivo. We establish a rat esophageal organoid (rEO) in vitro system and show that the landscapes of rEO formation, development and maturation trajectories can mimic those of rat esophageal epithelia in vivo. Single-cell RNA sequencing (scRNA-seq), snapshot immunostaining and functional analyses of stratified “matured” rEOs define that the epithelial pluripotent stem cell determinants, p63 and Sox2, play crucial but distinctive roles for regulating mammalian esophageal tissue homeostasis. We identify two cell populations, p63+Sox2+ and p63−Sox2+, of which the p63+Sox2+ population presented at the basal layer is the cells of origin required for esophageal epithelial stemness maintenance and proliferation, whereas the p63−Sox2+ population presented at the suprabasal layers is the cells of origin having a dual role for esophageal epithelial differentiation (differentiation-prone fate) and rapid tissue damage–repair responses (proliferation-prone fate). Given the fact that p63 and Sox2 are developmental lineage oncogenes and commonly overexpressed in ESCC tissues, p63−Sox2+ population could not be detected in organoids formed by esophageal squamous cell carcinoma (ESCC) cell lines. Taken together, these findings reveal that the tissue homeostasis is maintained distinctively by p63 and/or Sox2-dependent cell lineage populations required for the tissue renewing, damage–repair and protection of carcinogenesis in mammalian esophagi. [ABSTRACT FROM AUTHOR]

Human papillomavirus type 8 (HPV8), a cutaneous genus beta HPV type, has co-carcinogenic potential at sun-exposed sites in patients suffering from the inherited skin disease epidermodysplasia verruciformis (EV). We had previously shown that Langerhans cells responsible for epithelial immunosurveillance were strongly reduced at infected sites and that the HPV8 E7 protein interferes with the CCAAT/enhancer-binding protein (C/EBP)β to suppress the Langerhans cell chemokine CCL20. At the same time, however, we observed that EV lesions are heavily infiltrated with inflammatory immune cells, which is similar to the situation in HPV8 E6 transgenic mice. To identify critical inflammatory factors, we used a broad multiplex approach and found that the monocyte attracting chemokine CCL2 was significantly and strongly induced by HPV8 E6 but not E7-expressing HaCaT cells, which were used as a model for UV-damaged skin keratinocytes. Conditioned media from HPV8 E6-expressing keratinocytes enhanced CCL2-receptor (CCR2)-dependent monocyte recruitment in vitro, and macrophages predominated in the stroma but were also detected in the epidermal compartment of EV lesions in vivo. CCL2 induction by HPV8 E6 was even stronger than stimulation with the proinflammatory cytokine TNF-α, and both HPV8 E6 and TNF-α resulted in substantial suppression of the transcription factor C/EBPα. Using RNAi-mediated knockdown and overexpression approaches, we demonstrated a mechanistic role of the recently identified C/EBPα/miR-203/p63 pathway for HPV8 E6-mediated CCL2 induction at protein and transcriptional levels. Epithelial co-expression of p63 and CCL2 was confirmed in HPV8 E6-expressing organotypic air–liquid interface cultures and in lesional EV epidermis in vivo. In summary, our data demonstrate that HPV8 oncoproteins actively deregulate epidermal immune homeostasis through modulation of C/EBP factor-dependent pathways. While HPV8 E7 suppresses immunosurveillance required for viral persistence, the present study provides evidence that E6 involves the stemness-promoting factor p63 to support an inflammatory microenvironment that may fuel carcinogenesis in EV lesions.

A nuclear protein in testis (NUT) midline carcinoma arises from squamous cells and is often located in the head, neck, and lungs. This report focuses on the negative p63 mutation and older age at the diagnosis of a NUT carcinoma, which has significant prognostic implications. A 62-year-old patient presented initially with a three-year history of recurring frontal headaches, intermittent nasal bleeding, and a sensation of a nasal cavity mass. An incisional biopsy revealed a poorly differentiated NUT carcinoma in the left maxillary sinus. A functional endoscopic sinus surgery was performed, but the cancer recurred. As a result, a total maxillectomy was performed, and the patient was declared cancer-free with no evidence of residual disease. This is a rare instance of a p63-negative midline NUT cell carcinoma (NCC) in an elderly patient, which could potentially contribute to a more favourable prognosis and longer survival compared to other reported cases.

Respiratory viruses cause airway inflammation, resulting in epithelial injury and repair. miRNAs, including miR-149-5p, regulate different pathological conditions. We aimed to determine how miR-149-5p functions in regulating pro-inflammatory IL-6 and p63, key regulators of airway epithelial wound repair, in response to viral proteins in bronchial (BEAS-2B) and alveolar (A549) epithelial cells. BEAS-2B or A549 cells were incubated with poly (I:C, 0.5 µg/mL) for 48 h or SARS-CoV-2 spike protein-1 or 2 subunit (S1 or S2, 1 μg/mL) for 24 h. miR-149-5p was suppressed in BEAS-2B challenged with poly (I:C), correlating with IL-6 and p63 upregulation. miR-149-5p was down-regulated in A549 stimulated with poly (I:C); IL-6 expression increased, but p63 protein levels were undetectable. miR-149-5p remained unchanged in cells exposed to S1 or S2, while S1 transfection increased IL-6 expression in BEAS-2B cells. Ectopic over-expression of miR-149-5p in BEAS-2B cells suppressed IL-6 and p63 mRNA levels and inhibited poly (I:C)-induced IL-6 and p63 mRNA expressions. miR-149-5p directly suppressed IL-6 mRNA in BEAS-2B cells. Hence, BEAS-2B cells respond differently to poly (I:C), S1 or S2 compared to A549 cells. Thus, miR-149-5p dysregulation may be involved in poly (I:C)-stimulated but not S1- or S2-stimulated increased IL-6 production and p63 expression in BEAS-2B cells.

MicroRNAs (miRNAs) are small RNA molecules that regulate more than 30% of genes in humans. Recent studies have revealed that miRNAs play a crucial role in tumorigenesis. Large sets of miRNAs in human tumors are under-expressed compared to normal tissues. Furthermore, experiments have shown that interference with miRNA processing enhances tumorigenesis. Multiple studies have documented the causal role of miRNAs in cancer, and miRNA-based anticancer therapies are currently being developed. This review primarily focuses on two key points: (1) miRNAs and their role in human cancer and (2) the regulation of tumor suppressors by miRNAs. The review discusses (a) the regulation of the tumor suppressor p53 by miRNA, (b) the critical role of the miR-144/451 cluster in regulating the Itch-p63-Ago2 pathway, and (c) the regulation of PTEN by miRNAs. Future research and the perspectives of miRNA in cancer are also discussed. Understanding these pathways will open avenues for therapeutic interventions targeting miRNA regulation.

TP63 (p63) is strongly expressed in lower-grade carcinomas of the head and neck, skin, breast, and urothelium to maintain a well-differentiated phenotype. TP63 has two transcription start sites at exons 1 and 3′ that produce TAp63 and ΔNp63 isoforms, respectively. The major protein, ΔNp63α, epigenetically activates genes essential for epidermal/craniofacial differentiation, including ΔNp63 itself. To examine the specific role of weakly expressed TAp63, we disrupted exon 1 using CRISPR-Cas9 homology-directed repair in a head and neck squamous cell carcinoma (SCC) line. Surprisingly, TAp63 knockout cells having either monoallelic GFP cassette insertion paired with a frameshift deletion allele or biallelic GFP cassette insertion exhibited ΔNp63 silencing. Loss of keratinocyte-specific gene expression, switching of intermediate filament genes from KRT(s) to VIM, and suppression of cell-cell and cell-matrix adhesion components indicated the core events of epithelial-mesenchymal transition. Many of the positively and negatively affected genes, including ΔNp63, displayed local DNA methylation changes. Furthermore, ΔNp63 expression was partially rescued by transfection of the TAp63 knockout cells with TAp63α and application of DNA methyltransferase inhibitor zebularine. These results suggest that TAp63, a minor part of the TP63 gene, may be involved in the auto-activation mechanism of ΔNp63 by which the keratinocyte-specific epigenome is maintained in SCC.

The aim of the current study was to identify the expression of P63 and its relation to odontogenic epithelial cell proliferation, severity of the inflammatory infiltrate and size of radicular cysts (RCs). In this retrospective cross-sectional study, 30 cases of paraffin-embedded RCs were randomly selected from the archive. P63 and Ki-67 expression was assessed by immunohistochemistry. Epithelial P63 expression was absent in four (13.3%), weak in 10 (33.3%), and moderate in 16 (53.3%) cases. In the connective tissue wall of RC, P63 expression was absent in two (6.7%) cases, weak in 24 (80.0%) cases, and moderate in four (13.3%) cases. Ki-67 was found to be weakly expressed in 12 (40.0%) cases, moderately expressed in 13 (43.3%), and strongly expressed in five (16.7%) cases. No correlation was found between Ki-67 expression in odontogenic epithelium and P63 expression in the odontogenic epithelium (rho = 0.110, p =.563) or fibrous capsule (rho = 0.160, p =.399). Nevertheless, we found a positive correlation between Ki-67 expression in the odontogenic epithelium and the size of the RC (rho = 0.450, p =.013). The inflammatory infiltrate was negatively correlated with P63 expression in the odontogenic epithelium (rho = −0.428, p =.018), and with the size of cysts (rho = −0.728, p <.001). There is a high expression of P63 throughout the odontogenic epithelium and connective tissue capsule of the RC. P63 expression in the odontogenic epithelium is negatively correlated with the degree of the inflammatory infiltrate but not with epithelial cell proliferation or the size of the cyst. [ABSTRACT FROM AUTHOR]

Aim: To examine the expression of miR-203 and its target genes survivin and p63 in condyloma acuminatum (CA) and to determine their relationships with clinical recurrence. Methods: Case and control groups were assessed for general data and the expression levels of miR-203, survivin and p63. The case group received carbon dioxide laser treatment, had a 3-month follow-up period, and was then divided into recurrence and non-recurrence groups. The relevant data were analyzed using SPSS v. 25.0 software. Results:miR-203 was identified as an independent protective factor against the recurrence of CA, while wart number, survivin and p63 were independent risk factors for CA recurrence. Conclusion:miR-203 could be a specific clinical bioindicator for the prognosis and recurrence of CA. [ABSTRACT FROM AUTHOR]

Introduction: Amplification of human chromosome 3q26-29, which encodes oncoprotein ΔNp63 among other isoforms of the p63 family, is a feature common to squamous cell carcinomas (SCCs) of multiple tissue origins. Along with overexpression of ΔNp63, activation of the protooncogene, RAS, whether by overexpression or oncogenic mutation, is frequently observed in many cancers. In this study, analysis of transcriptome data from The Cancer Genome Atlas (TCGA) demonstrated that expression of TP63 mRNA, particularly ΔNp63 isoforms, and HRAS are significantly elevated in advanced squamous cell carcinomas of the head and neck (HNSCCs), suggesting pathological significance. However, how cooverexpressed ΔNp63 and HRAS affect the immunosuppressive tumor microenvironment (TME) is incompletely understood. Methods: Here, we established and characterized an immune competent mouse model using primary keratinocytes with retroviral-mediated overexpression of ΔNp63α and constitutively activated HRAS (v-rasHa G12R) to evaluate the role of these oncogenes in the immune TME. Results: In this model, orthotopic grafting of wildtype syngeneic keratinocytes expressing both v-rasHa and elevated levels of ΔNp63α consistently yield carcinomas in syngeneic hosts, while cells expressing v-rasHa alone yield predominantly papillomas. We found that polymorphonuclear (PMN) myeloid cells, experimentally validated to be immunosuppressive and thus representing myeloid-derived suppressor cells (PMN-MDSCs), were significantly recruited into the TME of carcinomas arising early following orthotopic grafting of ΔNp63α/v-rasHa-expressing keratinocytes. ΔNp63α/v-rasHa-driven carcinomas expressed higher levels of chemokines implicated in recruitment of MDSCs compared to vrasHa-initiated tumors, providing a heretofore undescribed link between ΔNp63α/HRAS-driven carcinomas and the development of an immunosuppressive TME. Conclusion: These results support the utilization of a genetic carcinogenesis model harboring specific genomic drivers of malignancy to study mechanisms underlying the development of local immunosuppression. [ABSTRACT FROM AUTHOR]

Aims: Due to its rarity and non‐specific clinical and pathological features, low‐grade adenosquamous carcinoma (LGASC) of the breast continues to pose diagnostic challenges. Unlike other triple‐negative breast carcinomas, LGASC tends to have an indolent clinical behaviour. It is essential to recognise this lesion for accurate diagnosis and appropriate management. Methods and results: Twenty‐five cases of LGASC were identified in our archives and collaborating institutes. Cases of LGASC with dominant coexisting other type carcinomas were excluded. We studied the clinical presentation, morphological features, patterns of the commonly used immunohistochemical stains and follow‐up. In our cohort, LGASC was commonly located at the outer aspect of the breast and associated with intraductal papilloma. The morphology of LGASC is characterised by infiltrating small glands and nests with variable squamous differentiation. We also found cuffing desmoplastic (fibrolamellar) stromal change in 75% of patients and peripheral lymphocytic aggregates in 87.5% of patients. P63 and smooth muscle myosin (SMM) were the most common myoepithelial markers used to assist in diagnosis. P63 often stained peripheral tumour cells surrounding invasive glands (circumferential staining in 80% of the cases), mimicking myoepithelial cells. It also stained the small nests with squamous differentiation. However, SMM was negative in 63% of the cases. The vast majority of our cases were triple‐negative; only a few had focal and weak expressions of ER and PR. One patient who did not have excision developed lymph node metastasis. Most patients underwent excision or mastectomy with negative margins as surgical treatment; there were no recurrences or metastases in these patients with clinical follow‐ups up to 108 months. Conclusions: LGASC has some unique, although not entirely specific, morphological features and immunohistochemical staining patterns. Fibrolamellar stromal change, peripheral lymphocytic aggregates and variable staining of p63 and SMM are valuable features to facilitate the diagnosis. [ABSTRACT FROM AUTHOR]

In men, prostate cancer (PCA) is the second most common malignancy and is the sixth leading cause of death amongst them. Differentiation of benign mimicker lesions (BML) of the prostate from PCA can be challenging sometimes, especially when limited amount of tissue is available. Several immuno-histochemical (IHC) markers are recommended for distinguishing such lesions; which becomes expensive for the patient. The present study was planned to study the utility of P63 IHC marker for diagnosis of benign, BML and adenocarcinoma of the prostate (ACP). The present - observational and analytical study - was conducted on 50 biopsy specimens of the prostate in the Department of Pathology at IIMSR, Lucknow during the period January 2017 to September 2022. The lesions included were: 20 specimens of benign prostatic hyperplasia (BPH), 17 of BML, 3 of prostatic intraepithelial neoplasia (PIN) and 10 of PCA. In the BML group, 5 specimens of basal cell hyperplasia (BCH), 4 specimens of simple atrophy (SA), 3 specimens of post atrophic hyperplasia (PAH) and 5 specimens of adenosis were studied. In the present study, all specimens of BPH and BMLare positive for P63 expression on IHC staining. In the PIN group, 2 out of 3 specimens were positive for P63. All 10 specimens of adenocarcinoma prostate were negative for P63 expression. We conclude that use of P63 IHC staining is adequate for distinguishing benign and BMLof prostate from ACPin majority of the specimens. [ABSTRACT FROM AUTHOR]

Background: The oral cavity is the most common site of cancer in the Indian subcontinent contributing to one-third of all cancers. The prognosis of oral dysplasia and malignant lesions can be predicted by both clinicopathological parameters and molecular markers. Aims and Objectives: The present study was undertaken to examine and compare the expression profile of cell cycle regulatory proteins p63 and Ki-67 in normal dysplastic and malignant lesions of oral mucosa and its correlation with prognosis. Settings and Design: This was a retrospective case-control study in a tertiary referral center. Materials and Methods: We evaluated and compared the expression profile of antigens p63 and Ki-67 on 60 cases of histologically proven oral dysplastic and malignant lesions oral squamous cell carcinoma (OSCC - 35 cases), oral epithelial dysplasia (OED - 15 cases), and 10 cases of benign and inflammatory oral lesions. Statistical Analysis: Mean and standard deviation were calculated and ANOVA statistical test was used to compare the data. Results: The mean labeling index of p63 and KI-67 was higher in OSCC than OED and minimum in inflammatory (controls) groups. Conclusion: p63 expression had a positive correlation with Ki-67 expression. Thus, p63 and Ki-67 may be a marker for tumorigenesis and tumor cell proliferation. [ABSTRACT FROM AUTHOR]

Primary cutaneous diffuse large B-cell lymphoma, leg type is a rare entity accounting for 4% of all primary cutaneous lymphomas whose clinical presentation encompasses a range of possibilities. COVID-19 has caused a delay in diagnosis of malignant neoplasms and consequently, this has resulted in poorer prognoses. A 62-year-old woman presented with two smooth-surfaced, mobile, well-circumscribed, oval, skin-colored nodules approximately one-cm in diameter with nonerythematous borders on the lower third of the left leg. Two months later, eleven nodules measuring between one and 1.5cm with erythematous halo, slight scaling, central erosion, and crusting had appeared. Histological study showed moderate pericapillary lymphocytic infiltration in the papillary and reticular dermis and prominent diffuse proliferation of medium to large cells in the subcutis. These exhibited irregular vesicular nuclei, a conspicuous solitary nucleolus of two to three small nucleoli, and three mitoses per high power field. Adipocytes were consistently encircled by neoplastic lymphocytes. Primary cutaneous diffuse large B-cell lymphoma, leg type is a high-grade lymphoma that can manifest as a diagnostic challenge and requires adequate immunohistochemistry and in situ hybridization studies for proper diagnosis, treatment, and prognosis.

The p53 family of tumor suppressors, which includes p53, p63, and p73, has a critical role in many biological processes, such as cell cycle arrest, apoptosis, and differentiation. In addition to tumor suppression, the p53 family proteins also participate in development, multiciliogenesis, and fertility, indicating these proteins have diverse roles. In this review, we strive to cover the relevant studies that demonstrate the roles of p53, p63, and p73 in lipid and iron metabolism.

Wei Ju,1,2,&ast; Guan-fa Luo,1,&ast; Yuan-yuan Shi,3 Fei-jun Zhou,1 Meng-qi Li,1 Jian-hui Xu,1 Zhi-xin Yan,2 Xi-hu Yang1 1Department of Oral and Maxillofacial Surgery, Affiliated Hospital of Jiangsu University, Zhenjiang, Jiangsu, 212001, People’s Republic of China; 2Department of Burn and Plastic Surgery, Affiliated Hospital of Jiangsu University, Zhenjiang, Jiangsu, 212001, People’s Republic of China; 3Department of Pathology, Affiliated Hospital of Jiangsu University, Zhenjiang, Jiangsu, 212001, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Xi-hu Yang, Email yangxihu1981@126.comObjective: Sebaceous carcinoma (SC) of the submandibular gland is extremely rare. Owing to the low morbidity and nonspecific clinical manifestations, diagnosis is commonly delayed, which increases metastasis and mortality. To date, there have been five reported cases of SC of the submandibular gland. Here, we present a new case and review the relevant literature.Methods and Results: A 36-year-old woman presented with an enlarged left submandibular gland. Clinical features included a non-tender solitary nodular mass with normal overlying skin. There were no special findings on computed tomography or ultrasound examination except for a swollen mass in the left submandibular gland. The patient underwent surgical resection. Pathological examination confirmed the diagnosis of SC with nerve infiltration. Immunohistochemical examination of this case showed positive staining for P63, P40, CK7, CK8/18, MLH1, MSH2, MSH6, and PMS2. The specimen was negative for androgen receptor, CEA, S-100, CK5/6, SOX-10, SOX-11, SMA, and GCDFP-15. The KI-67 labeling index was determined to be 15%. PAS and anti-epithelial membrane antigen were positive in partial area. The patient is still undergoing follow-up, and no metastasis or recurrence has been observed for 2 months.Conclusion: This case highlighted the fact that despite its rarity, SC should be considered as a differential diagnosis for masses located in the head and face. Early and accurate diagnosis, followed by wide surgical excision, has a favorable prognosis. Therefore, clinicians should be familiar with the clinical and pathological features of this disease.Keywords: sebaceous carcinoma, salivary gland neoplasms, submandibular gland, androgen receptor, P63, P40