Gene signatures in patients with early breast cancer and relapse despite pathologic complete response.

A substantial minority of early breast cancer (EBC) patients relapse despite their tumors achieving pathologic complete response (pCR) after neoadjuvant therapy. We compared gene expression (BC360; nCounter® platform; NanoString) between primary tumors of patients with post-pCR relapse (N = 14) with: (i) matched recurrent tumors from same patient (intraindividual analysis); and (ii) primary tumors from matched controls with pCR and no relapse (N = 41; interindividual analysis). Intraindividual analysis showed lower estrogen receptor signaling signature expression in recurrent tumors versus primaries (logFC = -0.595; P = 0.022). Recurrent tumors in patients with distant metastases also exhibited reduced expression of immune-related expression parameters. In interindividual analyses, primary tumor major histocompatibility complex class II expression was lower versus controls in patients with any relapse (logFC = -0.819; P = 0.030) or distant relapse (logFC = -1.151; P = 0.013). Primaries with later distant relapse also had greater homologous recombination deficiency than controls (logFC = 0.649; P = 0.026). Although no associations remained statistically significant following adjustment for false discovery rate, our results show that transcriptomic analyses have potential for prognostic value and may help in selecting optimal treatment regimens for EBC at risk of relapse and warrant further investigation.

Two progressed malignant phyllodes tumors of the breast harbor alterations in genes frequently involved in other advanced cancers.

BACKGROUND: The genomic landscape of phyllodes tumors (PTs) of the breast is not well defined, especially in patients with advanced disease. To shed light on this topic, paired primary and progressed tumor samples from two patients with malignant PTs were subjected to next-generation sequencing (NGS) followed by functional analysis of genetic alterations using two prediction tools. METHODS: The DNA of both the primary tumor and distant metastases of Patient 1 and the primary and recurrent tumor of Patient 2 were subjected to molecular profiling. NGS with the FoundationOne® assay was performed in a commercial molecular pathology laboratory. Two in silico prediction tools were used to estimate the pathogenicity of indicated genetic alterations. RESULTS: In total, 38 genomic alterations were detected, of which 11 were predicted to be probably benign. In Patient 1, 14 aberrations were identified in the primary tumor and 17 in pulmonary metastases, 12 of which were identical. In the primary and recurrent tumor of Patient 2, 17 and 15 sequence variants, respectively, were found, with 13 overlapping findings. Affected genes included seven (TP53, TERT, APC, ARID1A, EGFR, KMT2D, and RB1) of the top 10 most frequently altered genes in other advanced cancer entities, as well as four actionable therapeutic targets (EGFR, KIT, PDGFRA, and BRIP1). Of note, seven genes coding for receptor tyrosine kinases were affected: three in Patient 1 and four in Patient 2. Several genes (e.g. EPHA3, EPHA7, and EPHB1) were shown to be altered for the first time in PTs. CONCLUSIONS: The two progressed malignant PTs investigated here share some of the major genetic events occurring in other advanced cancers.

Granulomatous Mastitis: A Therapeutic and Diagnostic Challenge.

Granulomatous mastitis (GM) is a rare benign inflammatory breast disease that affects mostly women of childbearing age with a history of breastfeeding. The etiopathogenesis is still unknown; however, inflammation as the result of a reaction to trauma, metabolic or hormonal processes, autoimmunity, and an infection with Corynebacterium kroppenstedtii have all been implicated. Clinical findings are pain, mass, hyperemia, and inflammation. Because the clinical presentation can mimic infectious mastitis or inflammatory carcinoma, the disease course is often protracted. The diagnosis is made by histopathology. Biopsies show a granulomatous formation in combination with a localized infiltration of multi-nucleated giant cells, epithelioid histiocytes, and plasma cells. Ultrasound, mammography, and magnetic resonance imaging are not specific; however, ultrasound and mammography should be done to exclude other pathologies. Due to the lack of data including randomized controlled studies, the management of GM is controversial. In Western industrialized countries, most authors use a therapy regimen starting with antibiotics and corticosteroids, followed by continuous steroid therapy and surgery in patients with persisting symptoms. More data are needed to define the best therapy. The role of immunotherapy has not yet been ascertained. The implementation of a registry to collect more information on this rare disease is highly recommended.

The diagnostic utility of human papillomavirus-testing in combination with immunohistochemistry in advanced gynaecologic pelvic tumours: a new diagnostic approach.

Gynaecologic pelvic tumours comprise a range of histological entities that are highly variable with respect to their clinical behaviour. The distinction of these tumours can be extremely difficult. Accurate identification of the primary tumour site has significant impact on the treatment strategy and prognosis. In this study we investigated the diagnostic and clinical utility of HPV (Human Papillomavirus)-testing in combination with selected immunohistochemical markers in advanced pelvic tumours of unknown primary origin. Specimens of eight patients who presented with advanced gynaecologic pelvic tumour of unknown primary origin, 10 unequivocal cervical carcinomas with 10 corresponding metastases and 10 unequivocal endometrial carcinomas with 4 corresponding metastases were studied. All cases were analysed for HPV-infection by PCR. The expression of CEA, vimentin (VIM), estrogen receptor (ER) and progesterone receptor (PR) was studied by immunohistochemistry. HPV-DNA was exclusively detected in cervical carcinomas and their corresponding metastases but in none of the endometrial carcinomas (Fisher's exact test p<0.001). Endometrial carcinomas and their metastases were generally positive for ER (86%), PR (93%) and VIM (100%) but rarely positive for CEA (14%) and HPV (0%). Most cervical carcinomas and their corresponding metastases were positive for CEA (79%) and HPV (89%). They rarely expressed ER (5%), PR (5%) and VIM (10%). Among the eight patients with unknown primary tumour four patients were diagnosed with a cervical carcinoma and four with an endometrial carcinoma. In one case we could exclude an ovarian carcinoma. Immunohistochemical analysis of selected markers in combination with HPV-testing is simple and inexpensive. The detection of HPV-DNA seems to provide the most compelling evidence for the primary tumour site, when immunohistochemical analysis is less definitive. The proposed diagnostic approach is helpful in the identification and management of pelvic tumours of unknown primary origin.

Adenoviral expression of a transforming growth factor-beta1 antisense mRNA is effective in preventing liver fibrosis in bile-duct ligated rats.

BACKGROUND: Transforming growth factor-beta (TGF-beta) is a key mediator in establishing liver fibrosis. Therefore, TGF-beta as a causative agent may serve as a primary target for antifibrotic gene therapy approaches. We have previously shown that the adenoviral delivery of a transgene constitutively expressing a TGF-beta1 antisense mRNA blocks TGF-beta synthesis in culture-activated hepatic stellate cells and effectively abolishes ongoing fibrogenesis in vitro. METHODS: Ligature of the common bile duct was used to induce liver fibrosis in rats. The effect of the TGF-beta1 antisense on fibrogenesis was analyzed in this model of liver injury. RESULTS: In the present study, we demonstrate that the adenoviral vector directs the synthesis of mRNA quantities that are approximately 8000-fold more abundant than endogenous TGF-beta1 mRNA. In experimentally injured rat livers induced by ligature of the common bile duct, a model for persistent fibrogenesis and cirrhosis, administration of the adenoviral vector abrogates TGF-beta-enhanced production of collagen and alpha-smooth muscle actin. Furthermore, the number of cells positive for alpha-smooth muscle actin resulting from active recruitment of activated hepatic stellate cells around the bile ductular structures was significantly reduced in animals after application of Ad5-CMV-AS-TGF-beta1. However, the observed elevated serum levels of aspartate aminotransferase, alanine aminotransferase, and bilirubin induced in this obstructive liver injury model were not significantly altered in the presence of the TGF-beta antagonist. CONCLUSION: Taken together, our data provides in vivo evidence that the delivery of TGF-beta1 antisense mRNA specifically abolishes the diverse effects of direct TGF-beta function in ongoing liver fibrogenesis. Therefore, we conclude that the expressed transgene is therapeutically useful for inhibition of TGF-beta effects in diverse applications, ranging from clarification of TGF-beta function in the course of liver injury to the development of novel gene therapeutic approaches.

Malignant mediastinal germ cell tumor with pure hepatoid differentiation.

We describe a germ cell tumor of anterior mediastinal origin, with pure hepatoid differentiation and elevated serum AFP in a 41-year-old man. This is the first report of such a neoplasm analyzed by conventional stains and immunohistochemistry. Hepatocellular differentiation was proved by immunoreactivity with HepPar-1 and alpha-fetoproein (AFP), membranous expression of carcinoembryonic antigen (CEA-poly) in a canalicuar pattern, and focal expression of cytokeratin 19 in abortive ductular structures. Our investigation shows that mediastinal germ cell tumors with hepatoid components typically arise in middle-aged men; they are of pure hepatoid differentiation, as demonstrated here, or exclusively associated with yolk sac structures.