Joanna Szpor

Comprehensive immunohistochemical study of mesothelin (MSLN) using different monoclonal antibodies 5B2 and MN-1 in 1562 tumors with evaluation of its prognostic value in malignant pleural mesothelioma

Mesothelin (MSLN) is a glycophosphatidylinositol (GPI)-linked cell surface protein highly expressed in several types of malignant tumors sometimes in association with increased tumor aggressiveness and poor clinical outcome. In the present study, 1562 tumors were immunohistochemically analyzed for mesothelin expression using two different types of mouse monoclonal antibodies (5B2 and MN-1) to determine the clinical usefulness of mesothelin immunohistochemistry as well as to pinpoint potential targets for future anti-mesothelin therapy. Also, characterization of selected mesothelin-positive tumors was performed by immunohistochemistry and oncogene sequencing. Among the tumors analyzed, the highest frequencies of mesothelin-positivity were detected in ovarian serous carcinoma (90% in 5B2 and 94% in MN-1). Both antibodies showed frequent positivity in pancreatic adenocarcinoma (71% using 5B2 and 87% using MN-1) and malignant pleural mesothelioma (75% using 5B2 and 78% using MN-1). In malignant mesothelioma, overall survival was significantly longer in the cohort of patients with diffuse membranous expression of mesothelin (P < 0.001). Both antibodies showed positive staining in thymic carcinoma (77% in 5B2 and 59% in MN-1), however, no expression was detected in thymoma. No correlation was detected between mesothelin expression and mismatch repair system deficient phenotype or gene mutation (BRAF and RAS) status in gastrointestinal adenocarcinomas. Mesothelin immunohistochemistry may assist the differential diagnosis of thymoma vs. thymic carcinoma as well as prognostication of mesothelioma patients. Our results demonstrate that patients with solid tumors expressing mesothelin could be targeted by anti-mesothelin therapies.

Expression of ALCAM (CD166) and PD-L1 (CD274) independently predicts shorter survival in malignant pleural mesothelioma

Diffuse malignant mesothelioma of the pleura is a highly aggressive tumor typically associated with short survival. ALCAM (CD166), a type I transmembrane protein, is a member of the immunoglobulin superfamily. In normal cells, ALCAM regulates physiological processes such as angiogenesis and immune response. In cancer, it is associated with neoplastic progression, including invasion, migration, and metastasis. Furthermore, ALCAM is considered one of the cancer stem cell markers such as ALDH1 (ALDH1A1) and SALL4. The PD-L1 (CD274)/PD-1 (PDCD1, CD279) pathway is crucial for the modulation of immune responses in normal cells. Nevertheless, pathologic activation of the PD-L1/PD-1 pathway participates in immune evasion by tumor cells. Many PD-L1-expressing tumor cells have been identified in different types of cancer, including malignant mesothelioma. In this study, 175 well-characterized primary diffuse pleural mesotheliomas, including the epithelioid (n = 148), biphasic (n = 15), and sarcomatoid (n = 12) histotypes, were evaluated immunohistochemically for cancer stem cell markers (ALCAM, ALDH1, and SALL4) and PD-L1 expression. Twenty-five percent of the mesotheliomas (43/175) expressed ALCAM, whereas ALDH1 and SALL4 positivity was seen in 1% to 2% of cases. Thirty-three percent of the analyzed tumors (57/175) contained PD-L1-positive cells. Overall survival was significantly decreased in the cohort of patients with ALCAM- or PD-L1-positive tumors (both P < .01). Furthermore, the multivariate Cox hazards regression analysis identified ALCAM and PD-L1 (both P < 0.01) as potential independent risk factors. Thus, a combination of these 2 markers might be useful for prognostication and planning the treatment of patients with malignant pleural mesothelioma.

Prostate cancer with different ERG status may show different FOXP3-positive cell numbers

Prostatic carcinoma is the most frequent cancer in males in the Western world. A significant proportion of these cancers have a recurrent translocation involving ETS family genes, which leads to the overexpression of ERG transcription factor. Prostate cancers, which bear this mutation, differ in a number of features, including tumor microenvironment. One of the components of the tumor microenvironment is FOXP3 positive lymphocytes, which may participate in breaking immunosurveillance and promoting tumor growth. The aim of the study was to analyze the relationships between ERG expression, number of FOXP3 positive cells and other features of the tumor. The study group consisted of 65 cases. Tissue microarrays composed of 2 mm tissue cores were used for immunohistological evaluation. Immunohistochemistry for ERG and FOXP3 was performed according to the routinely applied protocol. The FOXP3 positive cells were counted and the results were expressed as the number of cells per mm2. The average number of FOXP3 positive cells was 33.30/mm2 for all cases, 21.43/mm2 for the ERG negative and 42.28/mm2 for the ERG positive group (p < 0.02). There were no significant relationships between FOXP3 positive cell count and any other parameters studied. Our results suggest that the immune response may differ between ERG negative and ERG positive prostatic carcinomas.

Recurrent vulvar melanoma in 35-year-old pregnant women.

Vulvar melanoma represents between 3% and 10% of vulvar neoplasms. We present a case of a 34-year-old pregnant woman presenting with a pigmented lesion on the left labium majus; she reported no family history of melanoma. The histological diagnosis was malignant melanoma, superficial spreading type, with Breslow thickness of 0.9 mm; the excision was complete. Eight months before, an atypical genital nevus was completely excised from a nearby location. The pregnancy was finished by cesarean delivery at term, and 3 months later, another pigmented lesion was noticed near but not within the scars. Partial right vulvectomy was performed, and histological diagnosis was malignant melanoma of superficial spreading type, with Breslow thickness of 0.7 mm. The specimen obtained in the first operation was reviewed, and although histological examination was diagnostic for atypical genital nevus, Vysis Melanoma Fluorescence in situ hybridization Probe Kit revealed increased copy numbers of RREB1, which could be consistent with a diagnosis of malignant melanoma.

CD207+/langerin positive dendritic cells in invasive and in situ cutaneous malignant melanoma

Introduction Dendritic cells are crucial for cutaneous immune response. Their role in melanoma progression is however a matter of controversy. Material and methods The number of dendritic cells within epidermis and in peri- and intratumoral location was analyzed using CD207 immunostain in 17 cases of in situ and 25 case of invasive melanoma. Results Average peritumoral CD207+ cells count was 22.88 for all cases, 17.94 for in situ lesions and 26.24 for invasive cases. Average epidermal CD207+ cells count was 164.47 for all cases, 183.00 for in situ lesions and 150.78 – for invasive cases. In case of invasive melanomas, peritumoral CD207+ cells count was positively correlated with Breslow stage (R = 0.59) mitotic activity within the tumor (R = 0.62). Invasive cases with regression showed higher intratumoral and epidermal CD207+ cells count than the ones without (275.00 vs. 95.32 and 173.20 vs. 148.35) but lower peritumoral CD207+ cells count (17.60 vs. 27.26). Invasive cases with ulceration showed higher intratumoral and peritumoral CD207+ cells count than the ones without ulceration (220.08 vs. 55.67 and 44.17 vs. 9.69). Conclusions CD207+ cells play a role in both progression and regression of melanoma but their exact role needs further studies.

Mast cells influence neoangiogenesis in prostatic cancer independently of ERG status

A significant proportion of prostatic adenocarcinomas show recurrent translocation leading to ERG expression. Previously we found that ERG+ cases have higher microvessel density than negative ones. One factor influencing angiogenesis in cancer is mast cells. The aim of the present study was to evaluate the relationship between microvessels, mast cells and ERG status. Tissue microarrays prepared from 113 radical prostatectomy specimens were analyzed with immunohistochemistry for CD31, tryptase and chymase. Vascular profiles and tryptase-positive and chymase-positive cells were counted. The average number of tryptase-positive cells was 28.93/mm2 and chymase-positive cells 9.91/mm2. The average number of CD31+ vascular profiles was 352.66/mm2. The average number of tryptase-positive cells was 26.35/mm2 for ERG- cases and 32.12/mm2 for ERG+ cases. The average number of chymase-positive cells was 8.14/mm2 for ERG- cases and 12.06/mm2 for ERG+ cases. The average number of CD31+ vascular profiles was 321.34/mm2 for ERG- cases and 390.74/mm2 for ERG+ cases. The number of CD31+ vascular profiles was positively correlated with the number of tryptase-positive and chymase-positive cells (R = 0.26 and R = 0.20). In summary, we demonstrated an interrelationship between mast cells, microvascular density and ERG status in prostatic carcinoma.

Application of tissue microarrays for receptor immunohistochemistry in breast carcinoma.

The current treatment of breast cancer, the most frequent malignancy found in females, requires the study of biomarkers. The standard set of these includes at least an estrogen receptor, a progesterone receptor and a HER2 receptor, although many other factors have been shown to contribute to the prognosis. Tissue microarrays have been introduced to decrease costs and workload of immunohistochemistry applied to large collections of samples. The aim of the study was to test the performance of this technology on three basic biomarkers of breast carcinoma in 106 cases of invasive breast carcinoma. Tissue microarrays composed of 3 cores sized 0.6 mm per case were constructed and stained by standard immunohistochemistry. The results were assessed on virtual slides created with an Aperio scanner. A sensitivity and specificity of 0.83 and 0.88 was obtained for the estrogen receptor, 0.76 and 0.88 for the progesterone receptor, 0.69 and 0.96 for HER2. In conclusion, TMA technology may give results comparable to the diagnosis based on whole sections, and the clinicopathologic correlations for the immunohistochemistry performed by both methods are fairy similar.

Clinical predictors of malignancy in patients diagnosed with atypical ductal hyperplasia on vacuum-assisted core needle biopsy

Introduction Atypical ductal hyperplasia (ADH) is a benign lesion, which due to the risk of coexisting cancer is classified as a lesion of uncertain malignant potential. Aim To identify clinical predictors of cancer underestimation in patients with ADH diagnosed after vacuum-assisted breast biopsy (VABB). Material and methods Between 2001 and 2016, a total of 3804 vacuum-assisted core needle biopsies were performed at the First Chair of General Surgery of the Jagiellonian University Medical College in Krakow, including 2907 ultrasound (US)-guided biopsies and 897 digital stereotactic procedures. Seventy-six women were diagnosed with ADH and 72 of them underwent subsequent surgical excision. Demographic factors, medical history, family history, clinical symptoms, type and size of lesion determined in imaging scans, size of biopsy needle, and presence of coexisting lesions in VABB specimens were analysed as potential predictors of malignancy underestimation. Results Underestimation of breast carcinoma occurred in 21 (29.2%) patients. The upgrade rate was significantly higher only in patients with a lesion visible both in mammography (MMG) and US examinations and combined BIRADS-5. Conclusions Vacuum-assisted core needle biopsy is a minimally invasive technique used in diagnosing ADH. As the risk of breast malignancy underestimation is relatively high, open surgical biopsy remains the recommended procedure, especially in patients with lesions detected both in mammography and US examination. As we could not identify the factors that preclude cancer underestimation, all the women diagnosed with ADH should be informed about the risk of cancer underestimation.

Melanomas and Dysplastic Nevi Differ in Epidermal CD1c

Background. Dendritic cells could be involved in immune surveillance of highly immunogenic tumors such as melanoma. Their role in the progression melanocytic nevi to melanoma is however a matter of controversy. Methods. The number of dendritic cells within epidermis, in peritumoral zone, and within the lesion was counted on slides immunohistochemically stained for CD1a, CD1c, DC-LAMP, and DC-SIGN in 21 of dysplastic nevi, 27 in situ melanomas, and 21 invasive melanomas. Results. We found a significant difference in the density of intraepidermal CD1c+ cells between the examined lesions; the mean CD1c cell count was 7.00/mm2 for invasive melanomas, 2.94 for in situ melanomas, and 13.35 for dysplastic nevi. The differences between dysplastic nevi and melanoma in situ as well as between dysplastic nevi and invasive melanoma were significant. There was no correlation in number of positively stained cells between epidermis and dermis. We did not observe any intraepidermal DC-LAMP+ cells neither in melanoma in situ nor in invasive melanoma as well as any intraepidermal DC-SIGN+ cells in dysplastic nevi. Conclusion. It was shown that the number of dendritic cells differs between dysplastic nevi, in situ melanomas, and invasive melanomas. This could eventually suggest their participation in the development of melanoma.

Leiomyosarcoma of inferior vena cava complicated by Budd-Chiari syndrome and disseminated intravascular coagulation — case report

Leiomyosarcoma of inferior vena cava is a rare malignant mesenchymal tumor of the venous system that typically occurs in adulthood. Correct and early recognition of leiomyosarcoma is very important, because a complete resection of the tumor (with occasionally chemio-or radiotherapy) can lead to prolonged survival. We report a case of a 54-year-old man suffering from the leiomyosarcoma of inferior vena cava with infiltration of retroperitoneum and right adrenal gland.