RESUMEN
Human papillomavirus (HPV) is an important causative factor of cervical cancer and is associated with nonsmall cell lung cancer (NSCLC). Merkel cell polyomavirus (MCPyV) is a rare and highly fatal cutaneous virus that can cause Merkel cell carcinoma (MCC). Although coinfection with oncogenic HPV and MCPyV may increase cancer risk, a definitive etiological link has not been established. Recently, genomic variation and genetic diversity in the MCPyV noncoding control region (NCCR) among ethnic groups has been reported. The current study aimed to provide accurate prevalence information on HPV and MCPyV infection/coinfection in NSCLC patients and to evaluate and confirm Korean MCPyV NCCR variant genotypes and sequences. DNA from 150 NSCLC tissues and 150 adjacent control tissues was assessed via polymerase chain reaction (PCR) targeting regions of the large T antigen (LT-ag), viral capsid protein 1 (VP1), and NCCR. MCPyV was detected in 22.7% (34 of 150) of NSCLC tissues and 8.0% (12 of 150) of adjacent tissues from Korean patients. The incidence rates of HPV with and without MCPyV were 26.5% (nine of 34) and 12.9% (15 of 116). The MCPyV NCCR genotype prevalence in Korean patients was 21.3% (32 of 150) for subtype I and 6% (nine of 150) for subtype IIc. Subtype I, a predominant East Asian strain containing 25 bp tandem repeats, was most common in the MCPyV NCCR data set. Our results confirm that coinfection with other tumor-associated viruses is not associated with NSCLC. Although the role of NCCR rearrangements in MCPyV infection remains unknown, future studies are warranted to determine the associations of MCPyV NCCR sequence rearrangements with specific diseases.
Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas , Variación Genética , Genotipo , Poliomavirus de Células de Merkel , Infecciones por Papillomavirus , Humanos , Carcinoma de Pulmón de Células no Pequeñas/virología , Carcinoma de Pulmón de Células no Pequeñas/genética , Femenino , Poliomavirus de Células de Merkel/genética , Poliomavirus de Células de Merkel/aislamiento & purificación , Persona de Mediana Edad , Masculino , Anciano , Infecciones por Papillomavirus/virología , Infecciones por Papillomavirus/complicaciones , Infecciones por Papillomavirus/epidemiología , República de Corea/epidemiología , Infecciones por Polyomavirus/virología , Infecciones por Polyomavirus/epidemiología , Infecciones por Polyomavirus/complicaciones , Papillomaviridae/genética , Papillomaviridae/clasificación , Adulto , Coinfección/virología , Coinfección/epidemiología , Neoplasias Pulmonares/virología , Anciano de 80 o más Años , Prevalencia , ADN Viral/genética , Infecciones Tumorales por Virus/virología , Infecciones Tumorales por Virus/complicaciones , Infecciones Tumorales por Virus/epidemiología , Reacción en Cadena de la Polimerasa , Virus del Papiloma HumanoRESUMEN
Knowledge of Human Polyomavirus (HPyV) infection in the anal area and its association with sexually transmitted infections such as Human Papillomavirus (HPV) and Human Immunodeficiency Virus (HIV) remains limited. Therefore, anal specimens from 150 individuals of both sexes were analyzed for screening purposes. HPV DNA was found in 50.7% of cases, with a predominance of high-risk (HR) genotypes. HPyV DNA was found in 39.3% of samples, with Merkel Cell Polyomavirus (MCPyV) being the most common, with a higher viral load than JCPyV and BKPyV. In addition, MCPyV viral load increased in people living with HIV (PLWH) with HPV infection (p < 0.0001).
Asunto(s)
Coinfección , Infecciones por VIH , Poliomavirus de Células de Merkel , Infecciones por Papillomavirus , Infecciones por Polyomavirus , Carga Viral , Humanos , Masculino , Femenino , Infecciones por VIH/virología , Infecciones por VIH/complicaciones , Infecciones por Papillomavirus/virología , Adulto , Persona de Mediana Edad , Coinfección/virología , Coinfección/epidemiología , Poliomavirus de Células de Merkel/genética , Poliomavirus de Células de Merkel/aislamiento & purificación , Infecciones por Polyomavirus/virología , Infecciones por Polyomavirus/epidemiología , ADN Viral/genética , Genotipo , Canal Anal/virología , Canal Anal/patología , Anciano , Adulto Joven , Papillomaviridae/genética , Papillomaviridae/aislamiento & purificación , Papillomaviridae/clasificación , Infecciones Tumorales por Virus/virología , Infecciones Tumorales por Virus/epidemiología , PrevalenciaRESUMEN
BACKGROUND: Merkel cell carcinoma (MCC) is an aggressive cancer with often poor outcomes. Limited biomarkers exist for predicting clinical outcomes. The Merkel cell polyomavirus (MCPyV) serum antibody test (AMERK) has shown potential for indicating better recurrence-free survival in a single-institution study. The study aimed to evaluate the link between initial AMERK serostatus and survival. Secondary objectives included examining the relationship between initial AMERK titer levels and tumor burden. METHODS: A retrospective cohort study across two institutions analyzed patients tested with AMERK within 90 days of MCC diagnosis. Regression models assessed the association of survival outcomes with serostatus, considering various factors. The relationship between AMERK titer and tumor burden indicators was evaluated using ANOVA. Significance testing was exploratory, without a fixed significance level. RESULTS: Of 261 MCC patients tested, 49.4% were initially seropositive (titer ≥75). Multivariable analysis showed that seropositivity improved recurrence, event-free, overall, and MCC-specific survival rates. Strong associations were found between initial AMERK titer and clinical, tumor, and nodal stages, tumor size, and disease extent. Notably, improved survival with seropositivity was observed only in patients with localized disease at initial presentation. CONCLUSION: Circulating antibodies to MCPyV oncoproteins, as indicated by the AMERK test, are linked with better survival in MCC patients with localized disease at presentation. This could enhance patient risk profiling and treatment personalization. The study's retrospective nature and exploratory analysis are key limitations. PLAIN LANGUAGE SUMMARY: Merkel cell carcinoma (MCC) is a potentially aggressive skin cancer, and tools to predict patient outcomes are limited. A blood test called anti-Merkel cell panel (AMERK), which checks for specific antibodies related to this cancer, might give us some clues. In this study, we looked at 261 MCC patients who took the AMERK test within 90 days of diagnosis. We found that patients with an initial positive AMERK result tended to have better outcomes, especially if their cancer was in the early stages. However, it is important to note that this study has limitations, including using retrospective data and exploratory analyses.
Asunto(s)
Anticuerpos Antivirales , Carcinoma de Células de Merkel , Poliomavirus de Células de Merkel , Neoplasias Cutáneas , Humanos , Carcinoma de Células de Merkel/virología , Carcinoma de Células de Merkel/sangre , Carcinoma de Células de Merkel/mortalidad , Carcinoma de Células de Merkel/diagnóstico , Carcinoma de Células de Merkel/inmunología , Poliomavirus de Células de Merkel/inmunología , Poliomavirus de Células de Merkel/aislamiento & purificación , Femenino , Masculino , Estudios Retrospectivos , Anciano , Pronóstico , Persona de Mediana Edad , Neoplasias Cutáneas/sangre , Neoplasias Cutáneas/virología , Neoplasias Cutáneas/mortalidad , Neoplasias Cutáneas/diagnóstico , Neoplasias Cutáneas/inmunología , Neoplasias Cutáneas/patología , Anticuerpos Antivirales/sangre , Anciano de 80 o más Años , Infecciones Tumorales por Virus/virología , Infecciones por Polyomavirus/sangre , Infecciones por Polyomavirus/diagnóstico , Infecciones por Polyomavirus/virología , Infecciones por Polyomavirus/inmunologíaRESUMEN
BACKGROUND: Merkel cell polyomavirus (MCPyV), a human polyomavirus that is unequivocally linked to merkel cell carcinoma (MCC), has been found in association with keratinocytes carcinomas (KC), especially basal cell carcinoma (BCC) and cutaneous squamous cell carcinoma (cSCC). Nevertheless, there is scarce information about the possible involvement of MCPyV in the development of KC. OBJECTIVES: To assess the presence of MCPyV DNA and Large-T Antigen (LT-Ag) via Polymerase Chain Reaction (PCR) and Immunohistochemistry (IHC) in cases of KC, and to correlate its presence with immunohistochemical markers p16, p53, and ki67, tumor type and subtype, sun-exposed location, and epidemiological data. METHODS: The prevalence of MCPyV DNA, LT-Ag, and immunohistochemical markers p16, p53, and ki67 was assessed by PCR and Immunohistochemistry (IHC) in 127 cases of KC, these results were correlated with tumor type and subtype, sun-exposed location, and epidemiological data. RESULTS: The MCPyV DNA was detected in 42.57% (43 of 101) cases by PCR, the LT-Ag was detected in 16.4% (20 of 122) of cases, p16 in 81.5% (97 of 119), p53 in 66.4% (83 of 125), ki67 in 89% (73 of 82). No correlation between MCPyV LT-Ag and DNA confronted with tumor type, subtype, location site, and immunohistochemical markers was found. A single correlation between the MCPyV LT-Ag and cSCC tumors and peri-tumoral lymphocyte cells was noted. STUDY LIMITATIONS: Further steps need to be taken to better evaluate the MCPyV influence and its possible role in KC carcinogenesis, as the evaluation of the virus genome state, the gene sequence that encodes LT-Ag in the KC tumor cells, and in situ hybridization for viral DNA or RNA in these cells. CONCLUSIONS: Despite the frequent detection of MCPyV in KC, the data available so far does not support the hypothesis of a causal relationship between them.
Asunto(s)
Antígenos Virales de Tumores , Biomarcadores de Tumor , Carcinoma de Células de Merkel , Carcinoma de Células Escamosas , Antígeno Ki-67 , Poliomavirus de Células de Merkel , Neoplasias Cutáneas , Proteína p53 Supresora de Tumor , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Antígenos Virales de Tumores/análisis , Biomarcadores de Tumor/análisis , Carcinoma Basocelular/virología , Carcinoma Basocelular/patología , Carcinoma de Células de Merkel/virología , Carcinoma de Células de Merkel/patología , Carcinoma de Células Escamosas/virología , Carcinoma de Células Escamosas/patología , Inhibidor p16 de la Quinasa Dependiente de Ciclina/análisis , ADN Viral/análisis , Inmunohistoquímica , Queratinocitos/virología , Queratinocitos/patología , Antígeno Ki-67/análisis , Poliomavirus de Células de Merkel/aislamiento & purificación , Reacción en Cadena de la Polimerasa , Infecciones por Polyomavirus/virología , Neoplasias Cutáneas/virología , Neoplasias Cutáneas/patología , Proteína p53 Supresora de Tumor/análisis , Infecciones Tumorales por Virus/virologíaAsunto(s)
Carcinoma de Células de Merkel , Neoplasias Cutáneas , Carcinoma de Células de Merkel/virología , Carcinoma de Células de Merkel/diagnóstico , Carcinoma de Células de Merkel/patología , Humanos , Neoplasias Cutáneas/virología , Neoplasias Cutáneas/diagnóstico , Neoplasias Cutáneas/patología , Poliomavirus de Células de Merkel/aislamiento & purificación , Poliomavirus de Células de Merkel/genéticaRESUMEN
BACKGROUND: Merkel cell carcinoma (MCC) is an aggressive malignant neuroendocrine tumour. There are two subsets of MCC, one related to Merkel cell polyomavirus (MCPyV) and the other to ultraviolet radiation (UVR). MCPyV-positive and MCPyV-negative MCCs have been considered to be different tumours, as the former harbour few DNA mutations and are not related to UVR, and the latter usually arise in sun-exposed areas and may be found in conjunction with other keratinocytic tumours, mostly squamous cell carcinomas. Two viral oncoproteins, large T antigen (LT; coded by MCPyV_gp3) and small T antigen (sT; coded by MCPyV_gp4), promote different carcinogenic pathways. OBJECTIVES: To determine which genes are differentially expressed in MCPyV-positive and MCPyV-negative MCC; to describe the mutational burden and the most frequently mutated genes in both MCC subtypes; and to identify the clinical and molecular factors that may be related to patient survival. METHODS: Ninety-two patients with a diagnosis of MCC were identified from the medical databases of participating centres. To study gene expression, a customized panel of 172 genes was developed. Gene expression profiling was performed with nCounter technology. For mutational studies, a customized panel of 26 genes was designed. Somatic single nucleotide variants (SNVs) were identified following the GATK Best Practices workflow for somatic mutations. RESULTS: The expression of LT enabled the series to be divided into two groups (LT positive, n = 55; LT negative, n = 37). Genes differentially expressed in LT-negative patients were related to epithelial differentiation, especially SOX9, or proliferation and the cell cycle (MYC, CDK6), among others. Congruently, LT displayed lower expression in SOX9-positive patients, and differentially expressed genes in SOX9-positive patients were related to epithelial/squamous differentiation. In LT-positive patients, the mean SNV frequency was 4.3; in LT-negative patients it was 10 (P = 0.03). On multivariate survival analysis, the expression of SNAI1 [hazard ratio (HR) 1.046, 95% confidence interval (CI) 1.007-1.086; P = 0.02] and CDK6 (HR 1.049, 95% CI 1.020-1.080; P = 0.001) were identified as risk factors. CONCLUSIONS: Tumours with weak LT expression tend to co-express genes related to squamous differentiation and the cell cycle, and to have a higher mutational burden. These findings are congruent with those of earlier studies.
Merkel cell carcinoma (MCC) is an aggressive form of skin tumour. There are two subtypes of MCC: one of them is related to a virus called Merkel cell polyomavirus (MCPyV); the other one is related to persistent exposure to sunlight. The aim of this research was to find differences between these subtypes in their molecular behaviour (the genes that are expressed and the mutations that may be found). To do this, we carried out two studies, one to investigate gene expression (the process cells use to convert the instructions in our DNA into a functional product such as a protein) and one to look at gene mutations (changes in the DNA sequence). We found that the tumours that were not related to MCPyV expressed genes related to epithelial differentiation (the process by which unspecialized cells gain features characteristics of epithelial cells, which, among other things, make up the outer surface of the body), which means that the origin of both MCC subtypes may be different. We also found that MCPyV-related tumours had fewer mutations. Our findings are important because they help us to understand the biology of the MCC subtypes and could help with the development of new treatments for people diagnosed with skin tumours.
Asunto(s)
Antígenos Virales de Tumores , Carcinoma de Células de Merkel , Poliomavirus de Células de Merkel , Infecciones por Polyomavirus , Factor de Transcripción SOX9 , Neoplasias Cutáneas , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Antígenos Virales de Tumores/genética , Carcinoma de Células de Merkel/virología , Carcinoma de Células de Merkel/genética , Carcinoma de Células de Merkel/patología , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Poliomavirus de Células de Merkel/genética , Poliomavirus de Células de Merkel/aislamiento & purificación , Mutación , Infecciones por Polyomavirus/genética , Infecciones por Polyomavirus/virología , Neoplasias Cutáneas/virología , Neoplasias Cutáneas/genética , Neoplasias Cutáneas/patología , Factor de Transcripción SOX9/genética , Infecciones Tumorales por Virus/genética , Infecciones Tumorales por Virus/virologíaRESUMEN
OBJECTIVE: Infection with human tumor viruses is one of the hypothesized causes of cancer. The current investigation aimed to explore the presence and quantitative analysis of a new human tumor virus, Merkel cell polyomavirus (MCPyV) in tissue samples of 114 patients with oral cavity lesions including oral squamous cell carcinoma (OSCC), oral lichen planus (OLP), Dysplasia and oral irritation fibroma (OIF) in Northern Iran. METHODS: From 114 formalin fixed paraffin embedded samples; 35 with SCC, 29 with OLP, 14 with dysplasia and 36 with OIF were cut, deparaffinized and DNA was extracted. Quantitative detection of MCPyV large T antigen was performed by absolute quantitative Real-Time PCR. RESULT: MCPyV DNA was detected in 30.6% (n: 11/36) of IF, 24.1% (n; 7/29) of OLP, 21.4% (n:3/14) of dysplasia and 20% (n;7/35) of OSCC samples. The mean MCPyV DNA copy number was 2.32×10-2 ± 3.97 ×10-2, 2.02×10-2 (SD=3.13×10-2), 2.69×10-4 (SD=2.51×10-4), and 2.56×10-4 (SD=6.73×10-4) per cell in OSCC, dysplasia and both of OLP and OIF samples, respectively (P=0.76). CONCLUSION: This study provides the first data from Iran regarding the presence of MCPyV genome in oral cavity lesions and oral cancer. These results also emphasize that MCPyV has an active role in the occurrence of oral lesions and progression to cancer. Further studies should be carried out to clarify the role of MCPyV in oral cavity lesions.
Asunto(s)
Poliomavirus de Células de Merkel/aislamiento & purificación , Enfermedades de la Boca/epidemiología , Neoplasias de la Boca/epidemiología , Infecciones por Polyomavirus/epidemiología , Infecciones Tumorales por Virus/epidemiología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Biopsia , Carcinoma de Células de Merkel/epidemiología , Carcinoma de Células de Merkel/virología , Niño , ADN Viral/análisis , Femenino , Fibroma/epidemiología , Fibroma/virología , Neoplasias de Cabeza y Cuello/epidemiología , Neoplasias de Cabeza y Cuello/virología , Humanos , Irán/epidemiología , Liquen Plano Oral/epidemiología , Liquen Plano Oral/virología , Masculino , Poliomavirus de Células de Merkel/genética , Persona de Mediana Edad , Boca/virología , Enfermedades de la Boca/virología , Neoplasias de la Boca/virología , Infecciones por Polyomavirus/virología , Prevalencia , Reacción en Cadena en Tiempo Real de la Polimerasa , Neoplasias Cutáneas/epidemiología , Neoplasias Cutáneas/virología , Carcinoma de Células Escamosas de Cabeza y Cuello/epidemiología , Carcinoma de Células Escamosas de Cabeza y Cuello/virología , Infecciones Tumorales por Virus/virología , Adulto JovenRESUMEN
Merkel cell polyomavirus (MCPyV) infects most people asymptomatically, but recent reports indicate that the virus may be related to carcinogenesis. This study aimed to evaluate the impact of MCPyV on the development of papillary thyroid cancer (PTC). Totally, 1057 samples, including 412 fresh biopsy samples (FBS) and 645 paraffin-embedded PTC biopsy samples (PEBS), and 1057 adjacent non-cancerous samples were assessed for the presence of MCPyV DNA and RNA. MCPyV DNA was positive in 215 (20.3%) of samples, including 126 (30.6%) in FBS and 89 (13.8%) in PEBS. In MCPyV-positive samples, the mean MCPyV copy number was higher in the patients with FBS (2.3 × 10-1 ± 0.5 × 10-1 copies/cell) compared to PEBS (0.7 × 10-4 ± 0.1 × 10-4 copies/cell) and adjacent non-PTC normal samples (0.3 × 10-5 ± 0.02 × 10-5 copies/cell), indicating a statistically significant difference (P < 0.001). The LT-Ag RNA expression was higher in FBS compared to PEBS, while VP1 gene transcript was not detected in any samples. Although our findings showed the presence of MCPyV in a subset of PTC Iranian patients, further research is required to confirm these findings.
Asunto(s)
ADN Viral/genética , Poliomavirus de Células de Merkel/genética , Infecciones por Polyomavirus/complicaciones , Cáncer Papilar Tiroideo/virología , Neoplasias de la Tiroides/virología , Infecciones Tumorales por Virus/complicaciones , Carga Viral , Adulto , Estudios de Casos y Controles , Estudios Transversales , ADN Viral/análisis , Femenino , Humanos , Irán/epidemiología , Masculino , Poliomavirus de Células de Merkel/aislamiento & purificación , Persona de Mediana Edad , Infecciones por Polyomavirus/virología , Cáncer Papilar Tiroideo/epidemiología , Neoplasias de la Tiroides/epidemiología , Infecciones Tumorales por Virus/virologíaRESUMEN
The human ocular surface hosts a paucibacterial resident microbiome and virome. The factors contributing to homeostasis of this mucosal community are presently unknown. To determine the impact of ocular enucleation and prosthesis placement on the ocular surface microbiome, we sampled conjunctival swabs from 20 anophthalmic and 20 fellow-eye intact conjunctiva. DNA was extracted and subjected to quantitative 16S rDNA PCR, biome representational karyotyping (BRiSK), and quantitative PCR (qPCR) confirmation of specific organisms. 16S ribosomal qPCR revealed equivalent bacterial loads between conditions. Biome representational in silico karyotyping (BRiSK) demonstrated comparable bacterial fauna between anophthalmic and intact conjunctiva. Both torque teno virus and Merkel cell polyoma virus (MCPyV) were detected frequently in healthy and anophthalmic conjunctiva. By qPCR, MCPyV was detected in 19/20 anophthalmic samples compared with 5/20 fellow eyes. MCPyV copy number averaged 891 copies/ng in anophthalmic conjunctiva compared with 193 copies/ng in fellow eyes (p < 0.001). These results suggest that enucleation and prosthesis placement affect the ocular surface flora, particularly for the resident virome. As MCPyV has been shown to be the etiologic cause of Merkel cell carcinoma, understanding the mechanisms by which the ocular surface regulates this virus may have clinical importance.
Asunto(s)
Anoftalmos/genética , Bacterias/aislamiento & purificación , Poliomavirus de Células de Merkel/aislamiento & purificación , Torque teno virus/aislamiento & purificación , Anoftalmos/microbiología , Anoftalmos/patología , Anoftalmos/virología , Bacterias/genética , Bacterias/patogenicidad , Conjuntiva/microbiología , Conjuntiva/patología , Conjuntiva/virología , ADN Ribosómico/genética , Femenino , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Masculino , Células de Merkel/microbiología , Células de Merkel/patología , Células de Merkel/virología , Poliomavirus de Células de Merkel/genética , Poliomavirus de Células de Merkel/patogenicidad , Persona de Mediana Edad , Torque teno virus/genética , Torque teno virus/patogenicidadRESUMEN
The association of immune markers and clinicopathologic features and patient outcome has not been extensively studied in Merkel cell carcinoma (MCC). We correlated tumoral PD-L1 and IDO1 expression, and intratumoral CD8+ and FoxP3+ lymphocytes count with clinicopathologic variables, Merkel cell polyomavirus (MCPyV) status, and patient outcomes in a series of 132 MCC. By univariate analyses, tumoral PD-L1 expression >1% and combined tumoral PD-L1 >1% and high intratumoral FoxP3+ lymphocyte count correlated with improved overall survival (OS) (p = 0.016, 0.0072), MCC-specific survival (MSS) (p = 0.019, 0.017), and progression-free survival (PFS) (p = 0.043, 0.004, respectively). High intratumoral CD8+ and FoxP3+ lymphocyte count correlated with longer MSS (p = 0.036) and improved PFS (p = 0.047), respectively. Ulceration correlated with worse OS and worse MSS. Age, male gender, and higher stage (3 and 4) significantly correlated with worse survival. MCPyV positivity correlated with immune response. By multivariate analyses, only ulceration and age remained as independent predictors of worse OS; gender and stage remained for shorter PFS. Tumoral PD-L1 expression and increased density of intratumoral CD8+ lymphocytes and FoxP+ lymphocytes may represent favorable prognosticators in a subset of MCCs. Tumoral PD-L1 expression correlated with intratumoral CD8+ and FoxP3+ lymphocytes, which is supportive of an adaptive immune response.
Asunto(s)
Antígeno B7-H1/biosíntesis , Linfocitos T CD8-positivos/inmunología , Carcinoma de Células de Merkel/mortalidad , Indolamina-Pirrol 2,3,-Dioxigenasa/biosíntesis , Linfocitos Infiltrantes de Tumor/inmunología , Proteínas de Neoplasias/biosíntesis , Neoplasias Cutáneas/mortalidad , Subgrupos de Linfocitos T/inmunología , Inmunidad Adaptativa , Factores de Edad , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor , Linfocitos T CD8-positivos/química , Carcinoma de Células de Merkel/química , Carcinoma de Células de Merkel/inmunología , Carcinoma de Células de Merkel/virología , Femenino , Factores de Transcripción Forkhead/análisis , Neoplasias de Cabeza y Cuello/química , Neoplasias de Cabeza y Cuello/inmunología , Neoplasias de Cabeza y Cuello/mortalidad , Neoplasias de Cabeza y Cuello/virología , Humanos , Estimación de Kaplan-Meier , Recuento de Linfocitos , Masculino , Poliomavirus de Células de Merkel/aislamiento & purificación , Persona de Mediana Edad , Análisis Multivariante , Estadificación de Neoplasias , Neoplasias Primarias Secundarias/química , Neoplasias Primarias Secundarias/inmunología , Neoplasias Primarias Secundarias/mortalidad , Neoplasias Primarias Secundarias/virología , Pronóstico , Supervivencia sin Progresión , Modelos de Riesgos Proporcionales , Factores Sexuales , Neoplasias Cutáneas/química , Neoplasias Cutáneas/inmunología , Neoplasias Cutáneas/virología , Úlcera Cutánea/etiología , Infecciones Tumorales por VirusRESUMEN
Because the incidence of Merkel cell carcinoma (MCC) has increased significantly during the last 10 years and it is recognized that Merkel cell polyomavirus (MCPyV) and ultraviolet (UV) radiation represent two different etiological inputs sharing clinical, histopathological, and prognostic similar features, although with different prognosis, this study investigated the detection of MCPyV in skin and lymph nodes with histological diagnosis of MCC. Formalin-fixed paraffin-embedded tissue (FFPE) were retrieved from archived specimens and MCPyV non-coding control region (NCCR) and viral capsid protein 1 (VP1) sequences were amplified and sequenced. Results provide an interesting observation concerning the discrepancy between the MCPyV DNA status in primary and metastatic sites: in fact, in all cases in which primary and metastatic lesions were investigated, MCPyV DNA was detected only in the primary lesions. Our data further support the "hit-and-run" theory, also proposed by other authors, and may lead to speculation that in some MCCs the virus is only necessary for the process of tumor initiation and that further mutations may render the tumor independent from the virus. Few point mutations were detected in the NCCR and only silent mutations were observed in the VP1 sequence compared to the MCPyV MCC350 isolate. To unequivocally establish a role of MCPyV in malignancies, additional well-controlled investigations are required, and larger cohorts should be examined.
Asunto(s)
Carcinoma de Células de Merkel/virología , Poliomavirus de Células de Merkel/genética , Poliomavirus de Células de Merkel/aislamiento & purificación , Infecciones por Polyomavirus/virología , Neoplasias Cutáneas/virología , Anciano , Anciano de 80 o más Años , Proteínas de la Cápside/genética , Carcinoma de Células de Merkel/patología , Análisis Mutacional de ADN , ADN Viral/análisis , Femenino , Humanos , Masculino , Pronóstico , Neoplasias Cutáneas/patologíaRESUMEN
Merkel cell carcinoma (MCC) is a rare cutaneous neuroendocrine carcinoma of uncertain origin. MCC incidence varies by country and has been increasing among white populations over the last three decades. MCC occurs most commonly in elderly men and typically arises on sun-exposed areas. It is associated with a high mortality rate due to rapid growth and significant metastatic potential. Clinical and histopathological diagnosis are challenging, but prompt recognition of the disease is imperative for a correct management. Several hypotheses have been proposed to define the cell of origin, which still remains controversial. The discovery of Merkel cell polyoma virus, identified in the majority of MCCs, led to the hypothesis of the existence of two tumour subtypes showing biological, clinico-pathological and prognostic differences. Significant interest is nowadays directed to characterize MCC genomic alterations and microRNA (miRNA) expression profiles. Current treatment strategies for MCC depend on staging, and typically consist of surgery, radiation therapy, chemotherapy and/or, more recently, immunotherapy. The aim of this review is to provide an updated overview of MCC with a special focus on clinico-pathological aspects, molecular-genetics and therapy.
Asunto(s)
Carcinoma de Células de Merkel/genética , Carcinoma de Células de Merkel/patología , Neoplasias Cutáneas/genética , Neoplasias Cutáneas/patología , Antineoplásicos/uso terapéutico , Carcinoma de Células de Merkel/terapia , Carcinoma de Células de Merkel/virología , Aberraciones Cromosómicas , Humanos , Poliomavirus de Células de Merkel/aislamiento & purificación , Mutación , Pronóstico , ARN Mensajero/genética , Neoplasias Cutáneas/terapiaRESUMEN
Merkel cell carcinoma has been a focus of active scientific investigation in recent years and new information on the topic has emerged. Although uncommon, this primary cutaneous neuroendocrine carcinoma, usually involving the head/neck of elderly individuals, has a poor prognosis. Within the past two decades, an increase in the incidence of the tumor and the discovery of its link to the Merkel cell polyomavirus have focused medical attention on the lesion. The resulting studies have improved our understanding of the biology of the neoplasm and contributed to clinical care. Specifically, two pathogenic subsets of the tumor have come to light, the majority due to Merkel cell polyomavirus and the minority caused by ultraviolet radiation-induced genetic damage. This dichotomy carries prognostic implications favoring the former subset. In addition, having capitalized on the known susceptibility of the tumor to immune influences, investigators have recently discovered its responsiveness to immune checkpoint inhibition. This revelation has constituted a therapeutic milestone at the clinical level. Herein we provide an overview of the topic, outline updates in the field and place an emphasis on dermatopathologic aspects of Merkel cell carcinoma.
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Carcinoma de Células de Merkel/diagnóstico , Carcinoma de Células de Merkel/genética , Carcinoma Neuroendocrino/patología , Neoplasias Cutáneas/patología , Rayos Ultravioleta/efectos adversos , Anciano , Anciano de 80 o más Años , Carcinoma de Células de Merkel/epidemiología , Carcinoma de Células de Merkel/virología , Diagnóstico Diferencial , Femenino , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Humanos , Inhibidores de Puntos de Control Inmunológico/uso terapéutico , Incidencia , Masculino , Proteínas de la Membrana/metabolismo , Poliomavirus de Células de Merkel/aislamiento & purificación , Infecciones por Polyomavirus/complicaciones , PronósticoAsunto(s)
Carcinoma de Células de Merkel/mortalidad , Núcleo Celular/patología , Infecciones por Polyomavirus/diagnóstico , Neoplasias Cutáneas/mortalidad , Piel/patología , Carcinoma de Células de Merkel/diagnóstico , Carcinoma de Células de Merkel/terapia , Carcinoma de Células de Merkel/virología , Quimioterapia Adyuvante/estadística & datos numéricos , Humanos , Estimación de Kaplan-Meier , Poliomavirus de Células de Merkel/aislamiento & purificación , Infecciones por Polyomavirus/mortalidad , Infecciones por Polyomavirus/patología , Infecciones por Polyomavirus/virología , Pronóstico , Piel/citología , Piel/virología , Neoplasias Cutáneas/diagnóstico , Neoplasias Cutáneas/terapia , Neoplasias Cutáneas/virologíaRESUMEN
Merkel cell carcinomas (MCCs) are rare, aggressive, cutaneous neuroendocrine tumours, approximately 80% of which are caused by the genomic integration of Merkel cell polyomavirus (MCPyV). MCPyV-positive MCCs carry poor prognosis in approximately 70% of cases, highlighting the need for greater understanding of the oncogenic mechanisms involved in pathogenesis, progression and post-therapeutic relapse, and translation into novel therapeutic strategies. In a previous pilot study, we reported a potential relationship between MCPyV gene expression and oncogenic alternative Δ exon 6-7 TrkAIII splicing in formalin-fixed paraffin-embedded (FFPE) MCC tissues from a 12-patient cohort of >90% MCPyV-positive MCCs, diagnosed at San Salvatore Hospital, L'Aquila, Italy, characterising a new MCC subgroup and unveiling a novel potential MCPyV oncogenic mechanism and therapeutic target. This, however, could not be fully verified due to poor RNA quality and difficulty in protein extraction from FFPE tissues. Here, therefore, we extend our previous observations to confirm the relationship between MCPyV and oncogenic alternative Δ exon 6-7 TrkAIII splicing in fresh, nonfixed, MCPyV-positive MCC metastasis by detecting sequence-verified RT-PCR products, including full-length Δ exon 6-7 TrkAIII, and by Western blot detection of a 100 kDa TrkA protein isoform of identical size to 100 kDa Δ exon 6-7 TrkAIII expressed by stable transfected SH-SY5Y cells. We also report that in three MCC patients submitted for multidisciplinary treatment, including locoregional chemotherapy, MCPyV large T-antigen mRNA expression, Δ exon 6-7 TrkAIII mRNA expression and intracellular indirect immunofluorescence (IF) TrkA and phosphorylation protein isoform(s) immunoreactivity in FFPE tissues were not reduced in postchemotherapeutic-relapsed MCCs compared to pretherapeutic MCCs, extending the possible roles of this novel potential MCPyV oncogenic mechanism from MCC pathogenesis to post-therapeutic relapse and progression. Detection of alternative Δ exon 6-7 TrkAIII splicing in MCC, therefore, not only characterises a new MCPyV-positive MCC subgroup and unveils a novel potential MCPyV oncogenic mechanism but also identifies patients who may benefit from inhibitors of MCPyV T-antigen and/or TrkAIII expression or clinically approved Trk kinase inhibitors such as larotrectinib or entrectinib, which are known to inhibit activated TrkA oncogenes and to elicit durable responses in TrkA-fusion oncogene-driven cancers, supporting the call for a large-scale multicentre clinical study.
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Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Carcinoma de Células de Merkel , Infecciones por Polyomavirus , Receptor trkA/genética , Neoplasias Cutáneas , Infecciones Tumorales por Virus , Anciano , Anciano de 80 o más Años , Empalme Alternativo/genética , Anticuerpos Monoclonales Humanizados/administración & dosificación , Anticuerpos Monoclonales Humanizados/efectos adversos , Carcinoma de Células de Merkel/diagnóstico , Carcinoma de Células de Merkel/genética , Carcinoma de Células de Merkel/mortalidad , Carcinoma de Células de Merkel/terapia , Transformación Celular Neoplásica/genética , Terapia Combinada , Vías de Administración de Medicamentos , Femenino , Humanos , Comunicación Interdisciplinaria , Italia/epidemiología , Masculino , Poliomavirus de Células de Merkel/aislamiento & purificación , Poliomavirus de Células de Merkel/fisiología , Persona de Mediana Edad , Técnicas de Diagnóstico Molecular , Mutación , Grupo de Atención al Paciente , Infecciones por Polyomavirus/diagnóstico , Infecciones por Polyomavirus/genética , Infecciones por Polyomavirus/mortalidad , Infecciones por Polyomavirus/terapia , Pronóstico , Neoplasias Cutáneas/diagnóstico , Neoplasias Cutáneas/genética , Neoplasias Cutáneas/mortalidad , Neoplasias Cutáneas/terapia , Análisis de Supervivencia , Infecciones Tumorales por Virus/diagnóstico , Infecciones Tumorales por Virus/genética , Infecciones Tumorales por Virus/mortalidad , Infecciones Tumorales por Virus/terapiaRESUMEN
INTRODUCTION: The presence of Merkel cell polyomavirus (MCPyV) was identified in Merkel cell carcinoma (MCC). However, there was sparse information on the link of other common nonmelanoma skin cancers - basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) - to MCPyV infection. The current study describes the phylogenetic information of MCPyV isolated from Iranian non-MCC (nonmelanoma skin cancers) focusing on tumorigenesis of mutations in large tumor (LT) antigen (LT-Ag) fragment. METHODS: Sixty patients with BCC and 20 patients with SCC were included in this study (48 males and 32 females; average age 65 years). The MCPyV-DNA copy number in positive samples was measured by quantitative real-time PCR. Then, mutational analysis of the MCPyV LT gene was carried out by direct sequencing. RESULTS: While MCPyV DNA was detected in 6 (10%) of 60 BCC samples, no viral genome was found in SCCs. There was no distinct association of MCPyV positivity with gender, age, or type of tumor (BCC or SCC) (p value >0.05). Quantitative real-time PCR revealed that the median number of viral DNA copies per cell was 0.7 in 6 MCPyV-positive BCC samples. Furthermore, full-length LT-Ag sequencing of positive samples indicated no stop codon or frameshift mutations compared to reference sequences. CONCLUSION: Considering the important role of the LT-Ag in the pathogenicity of MCPyV, non-synonymous mutations compared with the reference proteins triggered relevant amino acid substitutions. Overall, the results showed no tumor-associated mutations in the LT-Ag sequence of MCPyVs from positive samples.
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Antígenos Virales de Tumores/genética , Carcinoma de Células de Merkel/genética , Carcinoma de Células Escamosas/genética , Poliomavirus de Células de Merkel/genética , Anciano , Anciano de 80 o más Años , Carcinoma de Células de Merkel/virología , Carcinoma de Células Escamosas/virología , Análisis Mutacional de ADN , ADN Viral/genética , Femenino , Expresión Génica , Técnicas Genéticas , Humanos , Irán/epidemiología , Masculino , Poliomavirus de Células de Merkel/aislamiento & purificación , Persona de Mediana Edad , Mutación , Filogenia , Reacción en Cadena en Tiempo Real de la Polimerasa , Neoplasias CutáneasRESUMEN
Merkel cell polyomavirus (MCPyV) is a common human skin pathogen, shows high seroprevalence and is considered the etiologic agent of Merkel cell carcinoma. However, studies which detect MCPyV DNA in blood products may reveal the importance of this virus for the transfusion medicine. In this study we analyzed by viral metagenomics 36 plasma samples obtained from blood donors positive for the common blood transmitted infections from the city of Macapá (Brazilian Amazon). The generated raw data were were analyzed through a specific bioinformatics pipeline aimed at discovery of emerging viruses. The genomes of interest were analyzed phylogeographically and phylogenetically. MCPyV complete genome was recovered from one HBV-positive pool with high coverage (~ 223×) indicating acute viremia or reactivated infection. Interestingly, the phylogeographic position of the identified strain suggests its ancestry compared to MCPyV isolate from Colombian Amazon which hypothesizes that viral dissemination in the Amazon may have originated from Brazil. In conclusion, this study brings information for the genetic relationships of MCPyV isolated from blood donors from the Brazilian Amazon and demonstrates the possible phylogeographic behavior of our strain in relation to the other findings. We also demonstrated a strong evidence of viremic MCPyV phase in blood donations, however, more studies are necessary in order to understand the MCPyV impact on transfusion therapy.
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Donantes de Sangre , Genoma Viral , Genómica , Poliomavirus de Células de Merkel/clasificación , Poliomavirus de Células de Merkel/genética , Infecciones por Polyomavirus/diagnóstico , Infecciones por Polyomavirus/virología , Brasil , ADN Viral , Evolución Molecular , Genómica/métodos , Humanos , Poliomavirus de Células de Merkel/aislamiento & purificación , Metagenómica , Filogenia , Estudios Seroepidemiológicos , Infecciones Tumorales por VirusRESUMEN
BACKGROUND: Since limited data are available about the prevalence of Merkel cell polyomavirus (MCPyV) and the genetic variability of its noncoding control region (NCCR) in the context of immunosuppression, this study aimed to investigate the distribution of MCPyV in anatomical sites other than the skin and the behavior of NCCR among an HIV-1-positive population. METHODS: Urine, plasma, and rectal swabs specimens from a cohort of 66 HIV-1-positive patients were collected and subjected to quantitative real-time polymerase chain reaction (qPCR) for MCPyV DNA detection. MCPyV-positive samples were amplified by nested PCR targeting the NCCR, and NCCRs alignment was carried out to evaluate the occurrence of mutations and to identify putative binding sites for cellular factors. RESULTS: MCPyV DNA was detected in 10/66 urine, in 7/66 plasma, and in 23/66 rectal samples, with a median value of 5 × 102 copies/mL, 1.5 × 102 copies/mL, and 2.3 × 103 copies/mL, respectively. NCCR sequence analysis revealed a high degree of homology with the MCC350 reference strain in urine, whereas transitions, transversions, and single or double deletions were observed in plasma and rectal swabs. In these latter samples, representative GTT and GTTGA insertions were also observed. Search for putative binding sites of cellular transcription factors showed that in several strains, deletions, insertions, or single base substitutions altered the NCCR canonical configuration. CONCLUSIONS: Sequencing analysis revealed the presence of numerous mutations in the NCCR, including insertions and deletions. Whether these mutations may have an impact on the pathogenic features of the virus remains to be determined. qPCR measured on average a low viral load in the specimens analyzed, with the exception of those with the GTTGA insertion.
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Coinfección/virología , ADN Viral/genética , Infecciones por VIH/virología , VIH-1/fisiología , Poliomavirus de Células de Merkel/genética , Infecciones por Polyomavirus/virología , ARN no Traducido/genética , Adulto , Anciano , Secuencia de Bases , Estudios de Cohortes , Coinfección/genética , Coinfección/inmunología , Estudios Transversales , Femenino , Infecciones por VIH/genética , Infecciones por VIH/inmunología , VIH-1/genética , Humanos , Terapia de Inmunosupresión , Masculino , Poliomavirus de Células de Merkel/aislamiento & purificación , Poliomavirus de Células de Merkel/fisiología , Persona de Mediana Edad , Adulto JovenAsunto(s)
Carcinoma de Células de Merkel/diagnóstico , Ganglios Linfáticos/patología , Metástasis Linfática/diagnóstico , Neoplasias Primarias Desconocidas/diagnóstico , Neoplasias Cutáneas/patología , Biomarcadores de Tumor/análisis , Carcinoma de Células de Merkel/secundario , Carcinoma de Células de Merkel/virología , Humanos , Ganglios Linfáticos/virología , Metástasis Linfática/patología , Poliomavirus de Células de Merkel/aislamiento & purificación , Neoplasias Primarias Desconocidas/virología , Neoplasias Cutáneas/diagnóstico , Neoplasias Cutáneas/virologíaRESUMEN
The literature suggests that p63 expression in Merkel cell carcinoma (MCC) is associated with a poor prognosis. p63 immunohistochemistry marks the 2 main isoforms of this transcriptional protein: TAp63 (tumor suppressor-like properties) and ∆Np63 (oncogenic properties). Little information about the isoform of relevance in MCC exists. p40 immunohistochemistry specifically marks ∆Np63, and using comparative, semiquantitative expression of p63 and p40, we sought to clarify the issue. Our cohort of 53 cases (28 men and 25 women, median age 79 years, interquartile range 71-88) was stratified by morphology and viral status. Immunohistochemistry (p63, p40, and cytokeratin 5/6) was performed, H-scores for nuclear expression of p63 and p40 were derived (2 observers; positivity ≥ 10), and interobserver agreement was evaluated. Clinical, pathological, and outcome data were documented. The results were analyzed statistically. Mortality amounted to 57% (median follow-up 686 days, interquartile range 292-1599). Positivity for Merkel cell polyomavirus was observed in 29 (55%) of cases. Expression of p63 and p40 was present in 36 (69%) and 4 (8%) of cases, respectively. Increased age (P = .0241), negative Merkel cell polyomavirus status (P = .0185), and p63 positivity (P = .0012) were significantly associated with mortality. The latter 2 variables were highly correlated (P = .004). The interclass correlation between the 2 sets of H-scores was 0.95. Our findings support an association between p63 expression and reduced overall survival in MCC and show consistency in scoring this prognostic parameter. TAp63 is the dominant isoform of the protein involved. The paradoxical tumor suppressor-like activity of this isoform in p63-positive MCCs with reduced overall survival requires further study.