Olaparib monotherapy as primary treatment in unselected triple negative breast cancer.

BACKGROUND: The antitumor efficacy of PARP inhibitors (PARPi) for breast cancer patients harboring germline BRCA1/2 (gBRCA1/2) mutations is well established. While PARPi monotherapy was ineffective in patients with metastatic triple negative breast cancer (TNBC) wild type for BRCA1/2, we hypothesized that PARPi may be effective in primary TNBCs without previous chemotherapy exposure. PATIENTS AND METHODS: In the phase II PETREMAC trial, patients with primary TNBC >2 cm received olaparib for up to 10 weeks before chemotherapy. Tumor biopsies collected before and after olaparib underwent targeted DNA sequencing (360 genes) and BRCA1 methylation analyses. In addition, BRCAness (multiplex ligation-dependent probe amplification), PAM50 gene expression, RAD51 foci, tumor-infiltrating lymphocytes (TILs) and PD-L1 analyses were performed on pretreatment samples. RESULTS: The median pretreatment tumor diameter was 60 mm (range 25-112 mm). Eighteen out of 32 patients obtained an objective response (OR) to olaparib (56.3%). Somatic or germline mutations affecting homologous recombination (HR) were observed in 10/18 responders [OR 55.6%, 95% confidence interval (CI) 33.7-75.4] contrasting 1/14 non-responders (OR 7.1%; CI 1.3-31.5, P = 0.008). Among tumors without HR mutations, 6/8 responders versus 3/13 non-responders revealed BRCA1 hypermethylation (P = 0.03). Thus, 16/18 responders (88.9%, CI 67.2-96.9), in contrast to 4/14 non-responders (28.6%, CI 11.7-54.7, P = 0.0008), carried HR mutations and/or BRCA1 methylation. Excluding one gPALB2 and four gBRCA1/2 mutation carriers, 12/14 responders (85.7%, CI 60.1-96.0) versus 3/13 non-responders (23.1%, CI 8.2-50.3, P = 0.002) carried somatic HR mutations and/or BRCA1 methylation. In contrast to BRCAness signature or basal-like subtype, low RAD51 scores, high TIL or high PD-L1 expression all correlated to olaparib response. CONCLUSION: Olaparib yielded a high clinical response rate in treatment-naïve TNBCs revealing HR deficiency, beyond germline HR mutations. TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT02624973.

Massive blow-out craters formed by hydrate-controlled methane expulsion from the Arctic seafloor.

Widespread methane release from thawing Arctic gas hydrates is a major concern, yet the processes, sources, and fluxes involved remain unconstrained. We present geophysical data documenting a cluster of kilometer-wide craters and mounds from the Barents Sea floor associated with large-scale methane expulsion. Combined with ice sheet/gas hydrate modeling, our results indicate that during glaciation, natural gas migrated from underlying hydrocarbon reservoirs and was sequestered extensively as subglacial gas hydrates. Upon ice sheet retreat, methane from this hydrate reservoir concentrated in massive mounds before being abruptly released to form craters. We propose that these processes were likely widespread across past glaciated petroleum provinces and that they also provide an analog for the potential future destabilization of subglacial gas hydrate reservoirs beneath contemporary ice sheets.

Low Protein A20 in Minor Salivary Glands is Associated with Lymphoma in Primary Sjögren's Syndrome.

Patients with primary Sjögren's syndrome (pSS) have an increased risk of developing lymphomas, particularly the subtype mucosa-associated lymphoid tissue (MALT) lymphoma. Chronic antigen stimulation and increased activation of nuclear factor-κB (NF-κB) are important factors for the pathogenesis of MALT lymphomas. Protein A20 is an inhibitor of NF-κB. A recent study of pSS-associated MALT lymphomas identified potential functional abnormalities in the TNFAIP3 gene, which encodes protein A20. The present study aimed to assess protein A20 by immunohistochemistry (IHC) in minor salivary glands (MSGs) and lymphoma tissue sections of patients with pSS and investigate a potential association with lymphoma development. Protein A20 staining in lymphocytes was scored in four categories (0 = negative, 1 = weak, 2 = moderate and 3 = strong). For statistical purposes, these scores were simplified into negative (scores 0-1) and positive (scores 2-3). We investigated associations between protein A20-staining, focus scores, germinal centre (GC)-like structures and monoclonal B-cell infiltration in MSGs. MSG protein A20 staining was weaker in pSS patients with lymphomas than in those without lymphomas (P = 0.01). Weak protein A20 staining was also highly associated with a lack of GC formation (P < 0.01). Finally, weaker A20 staining was observed in the majority of pSS-associated MALT lymphoma tissues. In conclusion, we found absent or weak protein A20 immunoreactivity in MSGs of patients with pSS with lymphomas. This finding indicates that protein A20 downregulation in lymphocytes might be a mechanism underlying lymphoma genesis in patients with pSS.

Epidemiology and classification of gastroenteropancreatic neuroendocrine neoplasms using current coding criteria.

BACKGROUND: The lack of uniform criteria for coding of gastroenteropancreatic neuroendocrine neoplasia (GEP-NEN) has hampered previous epidemiological studies. The epidemiology of GEP-NEN was investigated in this study using currently available criteria. METHODS: All patients diagnosed with GEP-NEN between January 2003 and December 2013 in a well defined Norwegian population of approximately 350 000 people were included. Age- and sex-adjusted incidence rates were calculated. The current 2010 World Health Organization criteria, European Neuroendocrine Tumour Society classification and International Union Against Cancer (UICC) classification were used. RESULTS: A total of 204 patients (114 male, 55.9 per cent) were identified. The median age at diagnosis was 61 (range 10-94) years. The annual overall crude incidence was 5.83 per 100,000 inhabitants, with an increasing trend (P = 0.033). The most frequent location was small intestine (60 patients, 29.4 per cent) followed by appendix (48 patients, 23.5 per cent) and pancreas (33 patients, 16.2 per cent). Grade 1 tumours were more common in gastrointestinal (100 patients, 58.5 per cent) than in pancreatic (9 patients, 27 per cent) NEN. According to the UICC classification, 77 patients (37.7 per cent) had stage I, 17 patients (8.3 per cent) stage II, 37 patients (18.1 per cent) stage III and 70 patients (34.3 per cent) had stage IV disease. No patient with stage I disease had grade 3 tumours; advanced tumour grade increased with stage. CONCLUSION: A high crude incidence of GEP-NEN, at 5.83 per 100,000 inhabitants, was noted together with a significant increasing trend over time.

Short term results of complete (D3) vs. standard (D2) mesenteric excision in colon cancer shows improved outcome of complete mesenteric excision in patients with TNM stages I-II.

BACKGROUND: The aim of the present study was to investigate whether the new method of complete mesocolic excision (CME) with a high (apical) vascular tie (D3 resection) had an immediate effect compared with a conventional (standard) approach even in those patients without lymph node metastases. METHODS: A cohort of 189 consecutive patients with tumour-nodal-metastasis (TNM) stages I-II and a mean age of 73 years were operated on in the period from January 2007 to December 2008 in three community teaching hospitals. The CME approach (n = 89), used in hospital A, was compared to the standard technique used (n = 105) in two other hospitals, B and C. Lymph node yields from the specimens were used as a surrogate measure of radical resections. Outcome was analysed after a median follow-up of 50.2 months. RESULTS: In-hospital mortality rate was 2.8 % in the CME group and 8.6 % in the standard group. The 3-year overall survival (OS) in the CME group was 88.1 versus 79.0 % (p = 0.003) in the standard group, and the corresponding disease-free survival (DFS) was 82.1 versus 74.3 % (p = 0.026). Cancer-specific survival was 95.2 % in the CME group versus 90.5 % in the standard group (p = 0.067). Age, operative technique, and T category were significant in multiple Cox regressions of OS and DFS. CONCLUSIONS: Compared with the standard (D2) approach, introduction of CME surgical management of colon cancer resulted in a significant immediate improvement of 3-year survival for patients with TNM stage I-II tumours as assessed by OS and DFS.

Tyrosine-kinase mutations in c-KIT and PDGFR-alpha genes of imatinib naïve adult patients with gastrointestinal stromal tumours (GISTs) of the stomach and small intestine: relation to tumour-biological risk-profile and long-term outcome

BACKGROUND: The identification of activating mutations in either c-KIT cell surface growth factor receptor or platelet-derived growth factor receptor alpha (PDGFRA) has lead the way for the development of novel agents that selectively inhibit key molecular events in gastrointestinal stromal tumour (GIST) pathogenesis. The aim of this study was to investigate the role of c-KIT and PDGFRA gene mutations in primary resectable, imatinib naïve GISTs located in the stomach and small intestine. METHODS: All adult patients with GIST located in either stomach or small intestine who underwent surgical resection without prior imatinib (Glivec) treatment were included. DNA extraction and mutational analysis were performed. Mutational analyses were performed for c-KIT (exons 9, 11, 13, and 17) and the PDGFRA genes (exons 12, 14 and 18). Clinical and pathological parameters were analyzed in relation to the mutations in c-KIT and PDGFRA. RESULTS: A total of 38 patients who underwent surgery for GIST located in either the stomach (n = 24) or in the small intestines (n = 14) were included. Mutations were found in 31 of 38 (81.6 %) patients, with 24 (63.2 %) located in c-KIT and 7 (18.4 %) in the PDGRFA exons, respectively. Seven patients (18.4 %) were wildtype (WT). The most common mutation was in c-KIT exon 11. Incidentally found GISTs were significantly smaller (size >5 cm in 15 % for incidental vs. 71 % for symptomatic; OR of 13.4, 95 % CI 2.3-76.5; P = 0.001) and had lower mitotic rate (0 % for incidental vs. 44 % of the symptomatic; OR 0.52, 95 % CI 0.36-0.75; P = 0.005). Accordingly, the Fletcher grade was significantly better for incidental cases, with most having very low or low risk (85 %) in contrast to 19 of 25 (76 %) symptomatic cases showing moderate to high-risk features (OR 17.4, 95 % CI 2.98-101.7; P < 0.001). However, the distribution of c-KIT, PDGFRA and WT was not differently distributed between incidental and symptomatic GISTs. Long-term survival up to 25 years (median: 8 years) was best determined by Fletcher risk-score in the multivariate model (HR 14.1, 95 % CI 1.7-114.5; p = 0.013). CONCLUSIONS: Long-term survival in resected GISTs of the stomach and small intestine is best determined by Fletcher risk-score. Mitotic activity appears related to tumour size and young age at onset. Mutational status did not influence the clinical or tumour-specific features in this cohort (AU)

Thrombotic microangiopathy and the antiphospholipid syndrome.

The antiphospholipid syndrome (APS) comprises the association between vascular thrombosis, including microthrombosis, pregnancy morbidity and the coexistence of anti-phospholipid antibodies. Thrombotic microangiopathy (TMA) can be one of the manifestations of the APS and may involve any organ. This feature of the APS is probably less recognized by clinical doctors than venous thrombosis and recurrent abortions. This case report presents a patient who developed a widespread TMA with renal failure, gastric mucosa ulceration, urinary bladder ulcerations and a finger necrosis as part of the APS.

Proliferation accurately identifies the high-risk patients among small, low-grade, lymph node-negative invasive breast cancers.

BACKGROUND: The proliferation factor mitotic activity index (MAI) is the strongest prognosticator in lymph node-negative invasive breast cancer patients under age 71. The question remains, whether this also holds for 'favourable prognosis' subgroups. PATIENTS AND METHODS: The study was a multicentre prospective analysis of the MAI for recurrence-free survival and overall cancer-related survival of grade, MAI, and other prognosticators in 853 long-term follow-up, T1-3N0M0 breast cancer patients under 71 years. RESULTS: In all tumours together (N = 853), in grade 3 (n = 269), in tumours <1 cm all grades (n = 84), 1-2 cm, grades 1 + 2 (n = 300), and 2-3 cm, grades 1 + 2 (n = 124), the MAI is prognostically superior. Other features [grade, estrogen receptor (ER), diameter, and age] did not enhance its prognostic value except in grades 1 + 2 tumours 2-3 cm diameter with MAI <10, where ER has an additional prognostic value. CONCLUSIONS: In women <71 years with T1-3N0M0 small or low-grade invasive breast cancer usually not receiving systemic treatment, MAI > or =10 accurately identifies those at high risk. These high-risk patients should be considered for adjuvant systemic therapy.

Cytological, histological, and clinical correlations in intravesical Bacillus Calmette-Guerin immunotherapy.

Bacillus Calmette-Guerin (BCG), an attenuated form of mycobacterium, has been used extensively for the immunotherapy of bladder carcinoma. BCG induces a granulomatous response in the bladder wall. The cytological features induced by BCG-immunotherapy in urines and bladder washes have been described in only rare articles. We evaluated cytologic specimens from 50 patients who received intravesical BCG for bladder carcinoma. Forty-three patients received BCG therapy in a single course, averaging to 6.3 weeks per patient, whereas seven patients received an average of 9 weeks therapy with 2 to 3 interruptions. A total of 93 voided urines (avg, 1.9/patient), 57 bladder washes (avg, 1.1/patient) and 64 biopsies (avg, 1.3/patient) were obtained in the 2-year and 8-month retrospective review from these 50 patients. There were 30 responders and 20 nonresponders to BCG therapy. Positive cytology was predictive of recurrence: (cytologic-histologic correlation, 85%). Nine of 64 (14%) tissue biopsies revealed granulomatous inflammation, whereas only 2 of 150 cytological specimens (1.3%) exhibited specific inflammatory cells and epithelioid cells diagnostic of granulomas. Decoy cells appeared on the surface of the urothelium in four biopsies and in two cytological specimens. Based on this study, cytology appears to be adequate for the follow-up of BCG-treated patients for assessment of recurrence of carcinoma. Diagnostic features of granulomas were detected only in a smaller percentage of urine specimens, compared with biopsies. The presence of decoy cells in the urine and biopsies is an intriguing phenomenon that requires further investigation, especially in regard to the source of these cells and whether they are apoptotic in nature or whether they result from necrosis.

Amelogenin dosage compensation in carcinoma of colon, lung, liver and kidney, is not a marker of clonality in males.

The analysis of patterns of X-chromosome inactivation is becoming increasingly utilized as a marker of clonal composition of tissues from women. To date, however, no analogous system has been found for the study of clonality in tissue from men. In the current study, the methylation patterns for portions of the amelogenin genes are tested, which are encoded on both the X- and Y-chromosome (AMGX and AMGY). The polymerase chain reaction (PCR) was used to amplify portions of AMGX and AMGY from genomic DNA of carcinomas of the colon, lung, liver and kidney, as well as from matched normal somatic tissues. The amplification target included Alu I methylation sensitive restriction endonuclease sites as well as a 189 bp sequence which is present in AMGX but is absent in AMGY. Polymerase chain reaction amplification of AMGX and AMGY was successful using genomic DNA from both tumour and normal control tissue in 24 of the 26 cases. Pretreatment of genomic DNA with Alu I blocked amplification of AMGX in all cases from both normal tissue and tumour. This indicates that AMGX and AMGY undergo a non-random pattern of methylation in both normal tissues and in tumours, precluding their use as a marker of clonality. Methylation of Alu I sites in AMGY suggests that the amelogenin genes undergo dosage compensation, which raises the possibility that the expression of amelogenin is not restricted to the development of the tooth bud but may also play some other role in various tissues of the body.

[A retrospective study on Icelandic men diagnosed with non-seminomatous testicular cancer 1971-1995.].

INTRODUCTION: Survival of patients with testicular cancer has changed dramatically over the last two decades. This is mainly related to more successful chemotherapy, using combinations of drugs including cisplatinum. Therapy with cisplatinum was started in 1978 in Iceland. The survival of Icelandic men with non-seminoma testicular cancer, before and after this change in therapy, is not known. OBJECTIVE: Therefore a retrospective population-based study was carried out on all Icelandic males diagnosed with non-seminoma testicular cancer between 1971 and 1995. MATERIAL AND METHODS: Fifty-seven males with an average age of 29.1 years (range 17-52) were included in the study. Clinical information was obtained from the Icelandic Cancer Registry and hospital records. All specimens were reexamined by a pathologist and the modified staging system of Boden and Gibb was used for staging the disease. Crude survival was evaluated with the Kaplan-Meier method. RESULTS: Age standardized incidence for non-seminoma testicular cancer was 1.8 / 100,000 males per year for the whole period. Among the 57 patients, testicular swelling (93%) and pain (56%) were the most common symptoms at diagnosis. All 57 patients underwent orchiectomy, and 37 received chemotherapy as well. The most common histological type was embryonal carcinoma (44%) and average tumor diameter was 4.3 cm with a range of 1-12 cm. Tventy-six (51%) patients had stage I disease at diagnosis but 10 (17%) had stage IV. Crude five and 10 year survival for the whole group was 85% and 83%. From 1971 to 1977 the crude five year survival was 36% but 98% for the period 1978-1995. In December 1995 seven (64%) of 11 patients diagnosed between 1971-1977 have died of the disease. On the other hand only two patients (4%) diagnosed after 1977 have died as of december 1995. One because of acute myelogenic leukemia, nearly seven years after diagnosis of testis cancer. The other died of teratocarcinoma 12 months after diagnosis despite intensive chemotherapy including cisplatinum. CONCLUSION: Survival of patients with non-seminoma testicular cancer in Iceland has improved dramatically after the introduction of cisplatinum based chemotherapy in 1978. Of 46 patients diagnosed after 1977 only one (2%) has died because of the disease and median follow up was eight years. The incidence is low compared to other Western countries if Norway and Danmark are not included, were the incidence is much higher. Clinical presentation of the disease is similar between these countries.

Analysis of clonality in archival tissues by polymerase chain reaction amplification of PGK-1.

Clonality of archival formalin-fixed tissue sections was analyzed by polymerase chain reaction amplification of a portion of the X-linked phosphoglycerate kinase (PGK-1) gene. Amplification was successful in 29 of 36 cases of uterine endometrioid adenocarcinoma. Five of these cases, including both tumor and control tissue from the same patients, were heterozygous for the BstXI polymorphic site of the PGK-1-amplified product, permitting analysis of clonality. Pretreatment of the DNA with HpaII blocked amplification of one of the two PGK-1 alleles from four of five cases of tumor, indicating the clonal pattern of X chromosome inactivation in these cases. In contrast, in DNA from paired control tissues HpaII pretreatment had no effect, indicating a random pattern of X chromosome inactivation in normal tissue. One of the cases of endometrioid adenocarcinoma contained a high proportion (45%) of nontumor cells, precluding the determination of clonality. We conclude that polymerase chain reaction amplification can be used for the determination of the pattern of X chromosome inactivation in formalin-fixed tissue sections. Such an approach makes it feasible to include specimens from archival tissue collections in the analysis of clonality.

Tumours in Iceland. 13. Malignant tumours of the testis. A histological classification, epidemiological considerations and survival.

Data from 76 patients with malignant tumours of the testis registered with the Icelandic Cancer Registry during the period 1955-1984 were analyzed and classified histologically according to the WHO criteria. In 55 patients the tumours were of one histological type (72% of total). In the order of frequency, seminoma was diagnosed in 35 (46%), embryonal carcinoma in 14 (18%) and teratoma in 6 patients (8%). Twenty patients had tumours of more than one histological type (26%), i.e. a combination of embryonal carcinoma and teratoma (teratocarcinoma) in nine (12%) and combined tumours (seminoma component with other tumours) in seven (9%). Two patients had choriocarcinomas and three had yolk sac (endodermal sinus) tumours, both of them in combination with other neoplasms. Embryonal rhabdomyosarcoma was the only malignant non-germ cell tumour diagnosed. The age-adjusted incidence of malignant testis tumours increased by 48% from 1.6 per 100,000 per annum in 1955-1964 to 3.1 per 100,000 in 1975-1984. The increase affected both seminoma and non-seminomatous tumours (NSGCT). There has been an improvement in survival with time. Sixteen deaths were related to testis tumours and all occurred within four years of diagnosis. The incidence of testicular tumours in Iceland is intermediate between "high risk" and "low risk" countries.