Basal Cell Carcinoma and World War II-Era Cathode Ray Oscilloscope Exposure

There is a high prevalence of skin cancer in World War II servicemen stationed in the Pacific theater as a result of various risk factors such as exposure to ultraviolet radiation and genetic predisposition. We sought to describe whether a possible association exists between basal cell carcinoma (BCC) development and the use of high-voltage cathode ray tube (CRT) oscilloscopes manufactured around 1940 to 1955, which were a source of X-radiation. We present a case series of 9 men aged 65 to 93 years who presented with similar head and neck distributions of BCC and a history of extensive use of early CRT oscilloscopes during and shortly after the World War II era. The patients were interviewed and their medical records reviewed to determine CRT exposure times and BCC location, subtype, and treatment. Representative BCC histologic sections were reviewed. A total of 230 BCCs of the head and neck region were identified and mapped. Questionnaires determined a minimum exposure of 600 (range, 624-9600) hours within a 60-cm distance of early CRT screens in all patients. The average number of aggressive histologic subtypes was 23.5%. The average number of Mohs micrographic surgery layers required to obtain negative margins was 1.99 compared with 1.63 in the control group treated by the same Mohs micrographic surgeon (P \u3c. 002). This descriptive study is the first to suggest that ionizing radiation from early CRT oscilloscopes may be a factor in the development of multiple BCCs of the head and neck with increased subclinical spread

Wilms’ tumor 1 (WT1) antigen is overexpressed in Kaposi Sarcoma and is regulated by KSHV vFLIP

In people living with HIV, Kaposi Sarcoma (KS), a vascular neoplasm caused by KS herpesvirus (KSHV/HHV-8), remains one of the most common malignancies worldwide. Individuals living with HIV, receiving otherwise effective antiretroviral therapy, may present with extensive disease requiring chemotherapy. Hence, new therapeutic approaches are needed. The Wilms' tumor 1 (WT1) protein is overexpressed and associated with poor prognosis in several hematologic and solid malignancies and has shown promise as an immunotherapeutic target. We found that WT1 was overexpressed in >90% of a total 333 KS biopsies, as determined by immunohistochemistry and image analysis. Our largest cohort from ACTG, consisting of 294 cases was further analyzed demonstrating higher WT1 expression was associated with more advanced histopathologic subtypes. There was a positive correlation between the proportion of infected cells within KS tissues, assessed by expression of the KSHV-encoded latency-associated nuclear antigen (LANA), and WT1 positivity. Areas with high WT1 expression showed sparse T-cell infiltrates, consistent with an immune evasive tumor microenvironment. We show that major oncogenic isoforms of WT1 are overexpressed in primary KS tissue and observed WT1 upregulation upon de novo infection of endothelial cells with KSHV. KSHV latent viral FLICE-inhibitory protein (vFLIP) upregulated total and major isoforms of WT1, but upregulation was not seen after expression of mutant vFLIP that is unable to bind IKKƴ and induce NFκB. siRNA targeting of WT1 in latent KSHV infection resulted in decreased total cell number and pAKT, BCL2 and LANA protein expression. Finally, we show that ESK-1, a T cell receptor-like monoclonal antibody that recognizes WT1 peptides presented on MHC HLA-A0201, demonstrates increased binding to endothelial cells after KSHV infection or induction of vFLIP expression. We propose that oncogenic isoforms of WT1 are upregulated by KSHV to promote tumorigenesis and immunotherapy directed against WT1 may be an approach for KS treatment

Cardiofaciocutaneous syndrome (CFC) with congenital peripheral neuropathy and nonorganic malnutrition: An autopsy study

Many phenotypic manifestations have been reported in cardiofaciocutaneous (CFC) syndrome, but none, to date, are pathognomonic or obligatory. Previous histopathological studies reported findings in skin and hair; no autopsy studies have been published. We report the clinical and autopsy findings of a 7-year-old boy with severe CFC syndrome and malnutrition of psychosocial origin. Manifestations of CFC, reported previously, included macrocephaly and macrosomia at birth; short stature; hypotonia; global developmental delays; dry, sparse thin curly hair; sparse eyebrows and eyelashes; dilated cerebral ventricles; high cranial vault; bitemporal constriction; supraorbital ridge hypoplasia; hypertelorism; ptosis; exophthalmos; depressed nasal bridge; anteverted nostrils; low-set, posteriorly-rotated, large, thick ears; decayed, dysplastic teeth; strabismus; hyperelastic skin; wrinkled palms; keratosis pilaris atrophicans faciei; ulerythema ophryogenes; hyperkeratosis; gastroesophageal reflux; and tracheobronchomalacia. Additional findings, not previously reported, include islet cell hyperplasia, lymphoid depletion, thymic atrophy and congenital hypertrophy of peripheral nerves with onion bulb formations. Although the islet cell hyperplasia, lymphoid depletion, and thymic atrophy are nonspecific findings that may be associated with either CFC or malnutrition, the onion bulb hypertrophy is specific for a demyelinating-remyelinating neuropathy. These findings implicate congenital peripheral neuropathy in the pathogenesis of the developmental delays, feeding difficulties, respiratory difficulties, ptosis and short stature in this case. Additional studies of other cases of CFC are needed. (c) 2005 Wiley-Liss, Inc.Univ S Alabama, Womens & Childrens Hosp, Mobile, AL 36688 USAUniversidade Federal de São Paulo, São Paulo, BrazilUniversidade Federal de São Paulo, São Paulo, BrazilWeb of Scienc

vFLIP upregulates major oncogenic WT1 isoforms.

A. vFLIP expression upregulates WT1, demonstrated by immunofluorescence (green) in HuARLT-1 cells transduced with a doxycycline inducible pLVX vFLIP-FLAG lentivirus for wild type vFLIP vs mutant vFLIP compared to untransfected control cells. A mutant vFLIP that is unable to bind IKKγ and induce NFĸB is defective in its ability to upregulate WT1. B. Quantitation of WT1 immunofluorescence upon vFLIP induction, performed by averaging the GFP fluorescence for six distinct areas, showed increase WT1 expression with wild type vFLIP compared to control (HuARLT-1 not transduced with vFLIP), and mutant vFLIP: control vs. vFLIP. Statistical significance was determined using one way ANOVA, Tukey’s multiple comparisons, pC. Western blots show that WT1 protein is increased upon vFLIP-FLAG induction in HuARLT-1 cells. EMD Millipore WT1-NT antibody (clone 6F-H2) against WT1 was used. This increase was not seen to the same extent when the mutant vFLIP-FLAG is expressed, which is also less stable than the wild type form, as seen with antibodies to FLAG. Quantification of three independent experiments is shown below, p = 0.05(*), using one-sided, student’s t test. D. Inhibition of NFĸB signaling with BMS-345541 demonstrated reduction of WT1 expression, with quantification for WT1 in three independent experiments shown below. E. RT-qPCR for WT1 showed induction of WT1 mRNA with wild type vFLIP expression, which did not occur upon mutant vFLIP expression: control vs. vFLIP, pF. Major WT1 isoforms, A, B, C, D are all found to be upregulated by RT-qPCR for WT1 in vitro upon vFLIP induction in HuARLT-1 cells, p<0.05(*), p<0.01(**), p <0.001(***), p<0.0001(****), not significant (ns), using ordinary one way ANOVA, Tukey’s multiple comparisons test.</p

RT-qPCR Primers.

In people living with HIV, Kaposi Sarcoma (KS), a vascular neoplasm caused by the KS herpesvirus (KSHV/HHV-8), remains the most common malignancy worldwide. Individuals living with HIV, receiving otherwise effective antiretroviral therapy, may present with extensive disease requiring chemotherapy. Hence, new therapeutic approaches are needed. The Wilms’ tumor 1 (WT1) protein is overexpressed and associated with poor prognosis in several hematologic and solid malignancies and has shown promise as an immunotherapeutic target. We found that WT1 was overexpressed in >90% of a total 333 KS biopsies, as determined by immunohistochemistry and image analysis. Our largest cohort from ACTG, consisting of 294 cases was further analyzed demonstrating higher WT1 expression was associated with more advanced histopathologic subtypes. There was a positive correlation between the proportion of infected cells within KS tissues, assessed by expression of the KSHV-encoded latency-associated nuclear antigen (LANA), and WT1 positivity. Areas with high WT1 expression showed sparse T-cell infiltrates, consistent with an immune evasive tumor microenvironment. We show that major oncogenic isoforms of WT1 are overexpressed in primary KS tissue and observed WT1 upregulation upon de novo infection of endothelial cells with KSHV. KSHV latent viral FLICE-inhibitory protein (vFLIP) upregulated total and major isoforms of WT1, but upregulation was not seen after expression of mutant vFLIP that is unable to bind IKKƴ and induce NFĸB. siRNA targeting of WT1 in latent KSHV infection resulted in decreased total cell number and pAKT, BCL2 and LANA protein expression. Finally, we show that ESK-1, a T cell receptor–like monoclonal antibody that recognizes WT1 peptides presented on MHC HLA-A0201, shows increased binding to endothelial cells after KSHV infection or induction of vFLIP expression. We propose that oncogenic isoforms of WT1 are upregulated by KSHV to promote tumorigenesis and that immunotherapy directed against WT1 may be an approach for KS treatment.</div

WT1 and LANA expression correlate with decreased T cell infiltrates.

A. Using HALO analysis software of KS tumors from the AMC066/A5263 clinical trial, pseudocolor images were created for IHC of LANA, WT1, CD8, and CD4 cells. B. WT1 expression in all cases was inversely correlated with the number of CD8+T cells, using Spearman’s correlation test. Of note, melanin is noted along the epidermis and was excluded upon analysis of WT1 expression. C. Overall assessment of cellular populations according to histological subtype showed lower CD8 and CD4 T cells with advanced histological subtype, and the reverse for WT1 and LANA, p D. Clusters of CD8+ cells were seen at the periphery of nodular KS lesions, a representative case. E. Areas with numerous LANA+ cells were selected and quantified for CD8 positivity in a sequential tissue section, and vice versa using ordinary one-way ANOVA, Tukey’s multiple comparisons test. This spatial analysis was performed for WT1, LANA, CD4+T cells and CD8+T cells in areas of high LANA (intranodular areas) or high CD8 +T cells (along the periphery of nodular lesions), and lymphocytic infiltrates in corresponding regions for WT1/LANA/CD8/CD4 in plaques and patches. Ordinary one-way ANOVA multiple comparisons was performed as shown in S4 Table, p<0.05 (*), p <0.01 (**), p <0.001(***), p <0.0001(****), ns (not significant).</p

Schematic of a Proposed Model of WT1 Mediated Tumorigenesis in KS and WT1 ESK-1 Immunotherapy.

A diagram demonstrating KSHV infection contributing to vFLIP mediated upregulation of WT1 through NFĸB activation. WT1 upregulation in combination with KSHV, potentially impact the tumor microenvironment, contributing to an immunosuppressed state, in the suppression of T cells. WT1 directed immunotherapy such as TCRm mAb may aid in reversing the effects of WT1 mediated tumorigenesis through the targeting of KS spindle cells overexpressing WT1 through the induction of antibody-dependent cellular cytotoxicity (ADCC). (Image created with Biorender.com).</p

De novo KSHV infection in HuARLT-1 endothelial cells is latent.

HuARLT-1 endothelial cells assessed by flow cytometry for mCherry (indicative of lytic reactivation) and GFP flourescence (consistent with constitutive KSHV Infection) after de novo BAC-16 KSHV infection under conditions used for WT1 detection. No mCherry was detected at 48 hours, compared to approximately 5% mCherry positive cells after treatment with 1mM sodium butyrate, and 1.42% in untreated vs 8% treated cells (in the above example) at 72 hours post infection, p (TIF)</p