Laveniya Satgunaseelan

Programmed cell death-ligand 1 expression in oral squamous cell carcinoma is associated with an inflammatory phenotype

Phase 2 clinical trials utilising novel anti-PD1/PD-L1 antibodies are being conducted in oral cavity squamous cell carcinoma (OSCC) patients. However, data regarding PD-L1 expression in OSCC is limited. The aim of this study was to characterise the PD-L1 immunohistochemical expression in OSCC and its association with clinicopathological factors. Clinicopathological review of 217 patients with OSCC was performed, including quantifying tumour-infiltrating lymphocytes. Immunohistochemistry with PD-L1, CD4 and CD8 was performed. Forty (18.3%) cases showed PD-L1 expression. Expression was significantly more frequent in females (p=0.013), tongue/buccal mucosal SCCs (p=0.05), and in tumours with a high lymphocytic infiltrate (p>0.001). Intratumoural heterogeneity of PD-L1 expression was observed in 30% of the cases. PD-L1 expression was not significantly associated with disease-free (p=0.82) or overall survival (p=0.93). PD-L1 expression occurred in a significant minority of OSCC and can be heterogeneous. Frequent PD-L1 expression in OSCCs in females and in tumours with high lymphocytic infiltrate may assist in the selection of patients who may respond to anti-PD1/PD-L1 therapies.

p16 expression independent of human papillomavirus is associated with lower stage and longer disease-free survival in oral cavity squamous cell carcinoma.

There is limited information regarding the incidence of p16 expression, its association with human papillomavirus (HPV) and prognosis in oral cavity squamous cell carcinoma (OSCC). The role of p16 in OSCC is evaluated in 215 cases using tissue microarrays (TMAs). p16 immunohistochemistry and HPV in situ hybridisation were performed on TMAs following histopathology review of 215 patients with OSCC in the Sydney Head and Neck Cancer Institute database. Thirty-seven (17.2%) cases showed p16 expression without association with HPV. p16 expression significantly decreased with increasing pT category (p=0.002). p16 expression was associated with longer disease-specific survival on univariable analysis (p=0.044) but not on multivariable analysis adjusting for depth of invasion. Amongst patients receiving adjuvant radiotherapy, patients with p16 expression had significantly longer disease-free and overall survival. p16 expression was seen in early stage OSCCs and was associated with better survival following surgery and radiotherapy. While not an independent predictor of survival, p16 may mediate its effects by contributing to reduced proliferative capacity, leading to smaller tumour size and lower invasive potential.

p16 expression in cutaneous squamous cell carcinoma of the head and neck is not associated with integration of high risk HPV DNA or prognosis

Head and neck cutaneous squamous cell carcinoma (HNcSCC) can present with cervical metastases without an obvious primary. Immunohistochemistry for p16 is established as a surrogate marker of human papillomavirus (HPV) in oropharyngeal cancer. p16 expression in HNcSCC needs to be elucidated to determine its utility in predicting the primary site. The aim of this study was to evaluate the rate of p16 expression in HNcSCC and its association with prognostic factors and survival. p16 immunohistochemistry was performed on 166 patients with high risk HNcSCC (2000-2013) following histopathology review. Chromogenic in situ hybridisation (CISH) for HPV was performed. Fifty-three (31.9%) cases showed strong, diffuse nuclear and cytoplasmic p16 expression including 14 (41%) non-metastatic and 39 (29.5%) metastatic tumours (p=0.21). HPV CISH was negative in all cases. p16 expression significantly increased with poorer differentiation (p=0.033), but was not associated with size (p=0.30), depth of invasion (p=0.94), lymphovascular invasion (p=0.31), perineural invasion (p=0.69), keratinisation (p=0.99), number of involved nodes (p=0.64), extranodal extension (p=0.59) or survival. Nearly 32% of HNcSCCs, particularly poorly differentiated HNcSCCs, show p16 expression. A primary HNcSCC should be considered in p16 positive neck node metastases in regions with high prevalence of HNcSCC. p16 expression is not associated with improved survival in HNcSCC.

Atypical EWSR1 FISH signal patterns in bone and soft tissue tumours:– diagnostic experience with 135 cases

Recurrent Ewing sarcoma breakpoint region 1 (EWSR1) gene rearrangements characterize a select group of bone and soft tissue tumours. In our routine diagnostic practice with fluorescence in‐situ hybridization (FISH), we have occasionally observed EWSR1 gene rearrangements in tumours not associated classically with EWSR1 translocations. This study aimed to review our institutional experience of this phenomenon and also to highlight the occurrence of unusual EWSR1 FISH signals (i.e. 5′ centromeric region or 3′ telomeric region signals) that do not fulfil the published diagnostic criteria for rearrangements.

Melanoma diagnosis may be a pitfall for optical coherence tomography assessment of equivocal amelanotic or hypomelanotic skin lesions

Amelanotic and hypomelanotic skin lesions can be difficult to diagnose clinically and dermoscopically.1-3 Evidence is emerging that optical coherence tomography (OCT) can detect superficial basal cell carcinoma (sBCC) with good sensitivity (79-95.7%) and specificity (75.3-96%).4 Given that sBCC is a common problem4, and that it may be treated noninvasively5, the potential benefits from using OCT as an adjunct to clinical diagnosis are high. However, if OCT is used without histopathological confirmation in this setting, there is a risk that more clinically aggressive malignant pathology, such as amelanotic /hypomelanotic melanoma (AHM), may be misdiagnosed and left to progress if it is inadequately treated. Indeed, this research was prompted after experiencing a clinical case where a lesion had typical clinical-dermoscopic and OCT features for sBCC, but histopathology revealed an amelanotic melanoma. This article is protected by copyright. All rights reserved.

Deep sequencing of circulating exosomal microRNA allows non-invasive glioblastoma diagnosis

Exosomes are nano-sized extracellular vesicles released by many cells that contain molecules characteristic of their cell-of-origin, including microRNA. Exosomes released by glioblastoma cross the blood-brain-barrier into the peripheral circulation, and carry molecular cargo distinct to that of ‘free-circulating’ miRNA. In this pilot study, serum exosomal-microRNAs were isolated from glioblastoma (n=12) patients and analyzed using unbiased deep sequencing. Results were compared to sera from age- and gender-matched healthy controls, and to grades II-III (n=10) glioma patients. Significant differentially expressed microRNAs were identified, and the predictive power of individual and subsets of microRNAs were tested using univariate and multivariate analyses. Additional sera from glioblastoma patients (n=4) and independent sets of healthy (n=9) and non-glioma (n=10) controls were used to further test the specificity and predictive power of this unique exosomal-microRNA signature. Twenty-six microRNAs were differentially expressed in serum exosomes from glioblastoma patients’ relative to healthy controls. Random forest modeling and data partitioning selected seven miRNAs (miR-182-5p, miR-328-3p, miR-339-5p, miR-340-5p, miR-485-3p, miR-486-5p and miR-543) as the most stable for classifying glioblastoma. Strikingly, within this model, six iterations of these miRNA classifiers could distinguish glioblastoma patients from controls with perfect accuracy. The seven-miRNA panel was able to correctly classify all specimens in validation cohorts (n=23). Also identified were 23 dysregulated miRNAs in IDHMUT gliomas, a partially overlapping yet distinct signature of lower grade glioma. Serum exosomal-miRNA signatures can accurately diagnose glioblastoma preoperatively. miRNA signatures identified are distinct from previously reported ‘free-circulating’ miRNA studies in GBM patients, and appear to be superior.

A case of acitretin-induced haemorrhagic lesions in Darier disease

ment of acquired idiopathic partial hypohidrosis. J. Dermatol. 2006; 33: 265–7. 4. Ohshima Y, Yanagishita T, Ito K et al. Treatment of patients with acquired idiopathic generalized anhidrosis. Br. J. Dermatol. 2013; 168: 430–2. 5. Sano K, Asahina M, Uehara T et al. Degranulation and shrinkage of dark cells in eccrine glands and elevated serum carcinoembryonic antigen in patients with acquired idiopathic generalized anhidrosis. J. Eur. Acad. Dermatol. Venereol. 2017; 31: 2097–103.

FISH analysis of selected soft tissue tumors: Diagnostic experience in a tertiary center

Fluorescence in situ hybridization (FISH) is an important ancillary tool for the classification of bone/soft tissue (BST) tumors. The aim of this study was to evaluate the contribution of FISH to the final classification of common BST entities in the molecular pathology department of the Royal Prince Alfred Hospital (RPAH), which is one of the most important referral centers for the management of sarcomas in Australia.

Spindle cell variant of diffuse large B cell lymphoma occurring in the breast

Sir, Spindle cell morphology in tumours of the breast is not an uncommon occurrence, however in the context of lymphoma it is exceedingly rare. Whilst the sarcomatoid variant of anaplastic large cell lymphoma (ALCL) is relatively well characterised, less than 80 reports of B-cell lymphoma with spindle cell features have been described and none in the breast. The spindle cell variant of B-cell lymphomas has a predilection for extranodal sites, including the skin, female genital tract, oral cavity, paranasal sinuses, and cranium. Interestingly, most cutaneous cases of spindle cell B-cell lymphoma comprise follicular lymphomas, whilst all noncutaneous cases reported to date are diffuse large B-cell lymphomas (DLBCL) of germinal centre immunophenotype. We present the first report in the literature of a case of the spindle cell variant of DLBCL (sp-DLBCL) occurring in the breast, a potential diagnostic pitfall for confusion with a metaplastic carcinoma or other spindle cell breast neoplasm. A 28-year-old female presented to her general practitioner with a solid mass in her left breast for which she had a core biopsy. The biopsy showed an atypical spindle cell lesion arranged in fascicles and sheets, with an admixed inflammatory infiltrate of lymphocytes and eosinophils. The periphery of the lesion showed entrapped terminal duct lobular units, with focal necrosis. Immunohistochemistry showed weak positive staining for SMA, focal positive staining for Bcl-2 and CD34, with negative staining for cytokeratins, desmin, CD31, ER, PR, S100, EMA and ALK. A haematopoietic marker was not performed at this point. The initial diagnosis at core biopsy was that of an ‘atypical spindle cell lesion of the breast’. Following the core biopsy, the patient had a wide local excision of the left breast mass (65 36 32 mm, weighing 48 g), which macroscopically showed an ill-defined, pale fibrous lesion, 41 mm in maximal diameter. Again on histology, an atypical spindle cell lesion was identified (Fig. 1). Arranged in a haphazard storiform pattern were pleomorphic large spindle cells, with irregular elongated nuclei with vesicular chromatin and small nucleoli, and a moderate amount of pale eosinophilic cytoplasm with indistinct cell borders. No epithelial component was identified. Frequent mitotic figures were seen. The spindle cells were focally noted to merge with areas of cells with more typical centroblastic morphology. An underlying lymph node was not identified and no follicular dendritic cell networks were seen on immunohistochemistry for CD21 and CD35. Immunohistochemistry for non-haemopoietic markers including AE1/AE3, ER, PR, SMA, desmin, S100, CD34, Erg, and CD117 were all negative. By contrast, the atypical cells showed positive immunohistochemical staining for CD45, CD20, PAX5, CD10 and Bcl6, and negative immunohistochemical staining for MUM1, Bcl2, CD3, CD43, CD30, ALK, CD5, cyclin D1, EBER and TdT. The Ki-67 proliferative index was approximately 90%. Moderate numbers of CD3 positive T cells were noted within the lesion

Breast implant-associated squamous cell carcinoma – a rare long term complication

We present a case of a 58-year-old female, who presented with pain and induration of her left breast in 2014, following the third surgical revision of her breast implants initially inserted in 1985. On histopathological review, a high grade epithelial malignancy was seen confined to the perimeter of the implant capsule, reminiscent of an infiltrating ductal carcinoma of the breast. Keratinisation and intercellular bridges were identified and the malignant cells showed strong diffuse staining with CK5/6 and p63, supportive of a diagnosis of breast implant-associated squamous cell carcinoma (SCC). She subsequently underwent a completion mastectomy, which showed no evidence of residual malignancy. Two cases of breast implant-associated SCC have been documented in the early 1990s. This case highlights the importance of recognising this entity as a long term complication of breast implants, and as a potential diagnostic pitfall in implant-associated malignancy.