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Simple Summary: PD-L1 is a marker that helps determine a tumor's immune status and is used to select patients for immune therapy. Methylation modifies gene expression by adding a methyl group to DNA without affecting its sequence. In our study, we assessed and correlated PD-L1 and methylation status in pulmonary adenocarcinomas with high and negative PD-L1 expression. We investigated the pathobiological functions of the highest-ranking genes and promoters in both groups by searching genomic databases for their role in carcinomas. We observed distinct methylation patterns between PD-L1 high- and low-expressing tumors, indicating differences in their biological characteristics and tumor development. We conducted a pilot study to analyze the differential methylation status of 20 primary acinar adenocarcinomas of the lungs. These adenocarcinomas had to be wild type in mutation analysis and had either high (TPS > 50%; n = 10) or negative (TPS < 1%; n = 10) PD-L1 status to be integrated into our study. To examine the methylation of 866,895 specific sites, we utilized the Illumina Infinium EPIC bead chip array. Both hypermethylation and hypomethylation play significant roles in tumor development, progression, and metastasis. They also impact the formation of the tumor microenvironment, which plays a decisive role in tumor differentiation, epigenetics, dissemination, and immune evasion. The gained methylation patterns were correlated with PD-L1 expression. Our analysis has identified distinct methylation patterns in lung adenocarcinomas with high and negative PD-L1 expression. After analyzing the correlation between the methylation results of genes and promoters with their pathobiology, we found that tumors with high expression of PD-L1 tend to exhibit oncogenic effects through hypermethylation. On the other hand, tumors with negative PD-L1 expression show loss of their suppressor functions through hypomethylation. The suppressor functions of hypermethylated genes and promoters are ineffective compared to simultaneously activated dominant oncogenic mechanisms. The tumor microenvironment supports tumor growth in both groups. [ABSTRACT FROM AUTHOR]

Simple Summary: Glioblastomas are highly malignant brain tumors. Despite intensive research, there are no curative therapies available at the present time. Since Programmed Cell Death Ligand 1 (PD-L1) is a promising novel candidate in precision medicine, we here performed molecular analysis on glioblastomas with and without noteworthy PD-L1 expression. We found that there are severe molecular differences in glioblastomas depending on the PD-L1 state. An analysis of the top differences revealed post-transcriptional and RNA-associated pathways being altered. Targeting these altered pathways opens novel therapeutic approaches in the fight against brain cancer. Glioblastomas are the most frequent primary brain tumors in adults. They show highly malignant behavior and devastating outcomes. Since there are still no targeted therapies available, median survival remains in the range of 12 to 15 months for glioblastoma patients. Programmed Cell Death Ligand 1 (PD-L1) is a promising novel candidate in precision medicine. Here, we performed integrated epigenome-wide methylation profiling of 866,895 methylation-specific sites in 20 glioblastoma samples comparing PD-L1 high- (i.e., TPS (tumor proportion score) > 30%) and PD-L1 low-expressing glioblastomas (i.e., TPS < 10%). We found 12,597 significantly differentially methylated CpGs (DMCG) (Δβ ≥ 0.1 and p-value < 0.05) in PD-L1 high- compared with PD-L1 low-expressing glioblastomas. These DMCGs were annotated to 2546 tiling regions, 139 promoters, 107 genes, and 107 CpG islands. PD-L1 high-expressing glioblastomas showed hypomethylation in 68% of all DMCGs. Interestingly, the list of the top 100 significantly differentially methylated genes showed the enrichment of regulatory RNAs with 19 DMCGs in miRNA, snoRNAs, lincRNAs, and asRNAs. Gene Ontology analysis showed the enrichment of post-transcriptional and RNA-associated pathways in the hypermethylated gene regions. In summary, dissecting the methylomes depending on PD-L1 status revealed significant alterations in RNA regulation and novel molecular targets in glioblastomas. [ABSTRACT FROM AUTHOR]

Purpose: Gliomas are the most frequent primary brain tumors of adults. Despite intensive research, there are still no targeted therapies available. Here, we performed an integrated analysis of glioma and programmed cell death ligand 1 (PD-L1) in 90 samples including 58 glioma and 32 control brain tissues. Methods: To identify PD-L1 expression in glioma, we performed immunohistochemical analysis of PD-L1 tumor proportion score (TPS) using the clinically valid PD-L1 22C3 antibody on 90 samples including controls and WHO grade I–IV gliomas. Results: We found that PD-L1 is highly expressed in a subfraction of glioma cells. Analysis of PD-L1 levels in different glioma subtypes revealed a strong intertumoral variation of PD-L1 protein. Furthermore, we correlated PD-L1 expression with molecular glioma hallmarks such as MGMT-promoter methylation, IDH1/2 mutations, TERT promoter mutations and LOH1p/19q. Conclusion: In summary, we found that PD-L1 is highly expressed in a subfraction of glioma, indicating PD-L1 as a potential new marker in glioma assessment opening up novel therapeutic approaches. [ABSTRACT FROM AUTHOR]

Aims ALK FISH analysis is used as the reference standard to demonstrate ALK rearrangements, which qualify patients with pulmonary adenocarcinomas for therapy with ALK inhibitors. The aim of this study was to find screening ALK antibody clones with the best positive and best negative percentage agreement with ALK FISH. Methods and results Three hundred and three pulmonary adenocarcinomas were evaluated with ALK FISH and stained with five ALK antibody clones (5A4; D5F3; ALK1; ALK01; SP8) with standardized detection systems. D5F3 was additionally assessed using the OptiView enhanced detection and amplification system. ALK FISH found 14 cases (4.6%) that harboured ALK rearrangements. These stained at all intensities for D5F3 and 5A4. To identify rearranged cases among stained cases, we subsequently analysed all immunohistochemically positive cases with ALK FISH. Conclusions D5F3 with OptiView exclusively stained rearranged cases with strong intensity, without a single false-positive or false-negative case. The number of subsequent ALK FISH analyses required would have decreased from 303 to 14 cases (−95.4%), reducing significantly the time, work and costs without any loss of diagnostic quality and accuracy. [ABSTRACT FROM AUTHOR]

We investigated stereoscopic imaging for gross examination in telepathology. A conventional macroscopic station was equipped with two cameras mounted 6.5 cm apart and images were produced of 30 different routine pathology specimens. Still images were displayed on a three-dimensional auto-stereoscopic display with a lenticular plate (which did not require the viewer to wear special glasses) and as a three-dimensional projection that required the viewer to wear glasses with polarized lenses. Nine observers (pathologists, laboratory technicians and engineers) viewed the three-dimensional images first on the auto-stereoscopic display and then with polarized projection. The observers scored the images for spatial reproduction, surface structure, proportions, colour and sharpness (10 indices in total, each rated on a five-point Likert scale of 1–5, with lower scores indicating better quality). Results were compared with those from five observers who had previously viewed the corresponding two-dimensional images on a conventional (two-dimensional) display. The mean scores across each of the 10 indices were 2.9 (two-dimensional display), 2.1 (auto-stereoscopic display) and 1.6 (polarized projection). All observers stated that the polarized projection had superior image quality with regard to resolution, colour and surface structures. The results obtained in the present study with still images have encouraged us to integrate stereoscopy into a dynamic telepathology system. [ABSTRACT FROM AUTHOR]

We evaluated a dynamic telepathology system in routine use after a two-year developmental phase. In two three-month studies, two years apart, all intraoperative frozen sections were examined by one pathologist via the telepathology system. Laboratory technicians sampled tissues at gross examination and were connected with the pathologist by videoconference. Specimens were immediately reviewed after the telepathology diagnosis by the same diagnostician. In the second study there were 342 cases. The mean time required to diagnose small specimens such as biopsies was 8 min (range 4–14); for specimens with extensive gross examination it was 22 min (range 10–45). The telepathology and traditional frozen-section diagnoses agreed in 99.4% of cases, but telepathology took two to four times longer for gross and histological examinations, and up to 10 times longer for histological examination only. There were five false diagnoses (1.5%), two of which originated from telepathology (0.6%) and were recognized in the subsequent light microscopy review. Telepathology is not a replacement for light microscopy, but should be regarded as a complementary technique. [ABSTRACT FROM AUTHOR]

: ObjectivesTo assess the value of transition zone and lateral sextant biopsies for the detection of prostate cancer after a previous sextant biopsy was negative.: MethodsA total of 74 prostates after radical prostatectomy were used to perform biopsies ex vivo. First, a sextant biopsy was taken, then two different rebiopsy techniques were performed. Rebiopsy technique A consisted of a laterally placed sextant biopsy and two cores per side of the transition zones only. Rebiopsy technique B included a standard sextant biopsy and two cores per side from the lateral areas of the prostate. The biopsies were taken using ultrasound guidance to sample the areas of interest precisely.: ResultsThe initial sextant biopsy found 39 prostate cancers. Rebiopsy technique A found 12 cancers (34%). In this group, a laterally placed sextant biopsy found 12 cancers; transition zone biopsies revealed cancer in 5 cases, but no additional tumor was found. Rebiopsy technique B detected 23 prostate cancers (66%). Fourteen tumors were found after a second standard sextant biopsy, and nine additional tumors were found in the lateral areas.: ConclusionsSextant biopsy has a low sensitivity of only 53%. A biopsy including the transition zones is not the ideal technique for detecting the remaining tumors. Therefore, transition zone biopsies should be reserved for patients with multiple previous negative biopsies of the peripheral zone. A subsequent sextant biopsy with additional cores from the lateral areas of the prostate is favorable if rebiopsy is necessary after a negative sextant biopsy. [Copyright &y& Elsevier]

Simple Summary: Glioblastoma is the most malignant brain tumor. To date, there is no curative therapy available. Since EGFR is an interesting candidate in precision medicine, we performed an integrated molecular analysis in glioblastoma with and without EGFR amplification. We found that there are significant molecular differences in glioblastoma, depending on the EGFR amplification state. Analysis of top differences revealed DNA replication and packaging, and chromatin and gene silencing pathways. Targeting these altered pathways may open novel targets in precision medicine. Glioblastoma IDH wildtype is the most frequent brain tumor in adults. It shows a highly malignant behavior and devastating outcomes. To date, there is still no targeted therapy available; thus, patients' median survival is limited to 12–15 months. Epithelial growth factor receptor (EGFR) is an interesting targetable candidate in advanced precision medicine for brain tumor patients. In this study, we performed integrated epigenome-wide DNA-methylation profiling of 866,895 methylation specific sites in 50 glioblastoma IDH wildtype samples, comparing EGFR amplified and non-amplified glioblastomas. We found 9849 significantly differentially methylated CpGs (DMCGs) with Δβ ≥ 0.1 and p-value < 0.05 in EGFR amplified, compared to EGFR non-amplified glioblastomas. Of these DMCGs, 2380 were annotated with tiling (2090), promoter (117), gene (69) and CpG islands (104); 7460 are located at other loci. Interestingly, the list of differentially methylated genes allocated eleven functionally relevant RNAs: five miRNAs (miR1180, miR1255B1, miR126, miR128-2, miR3125), two long non-coding RNAs (LINC00474, LINC01091), and four antisense RNAs (EPN2-AS1, MNX1-AS2, NKX2-2-AS1, WWTR1-AS1). Gene ontology (GO) analysis showed enrichment of "DNA replication-dependent nucleosome assembly", "chromatin silencing at rDNA", "regulation of gene silencing by miRNA", "DNA packaging", "posttranscriptional gene silencing", "gene silencing by RNA", "negative regulation of gene expression, epigenetic", "regulation of gene silencing", "protein-DNA complex subunit organization", and "DNA replication-independent nucleosome organization" pathways being hypomethylated in EGFR amplified glioblastomas. In summary, dissecting the methylomes of EGFR amplified and non-amplified glioblastomas revealed altered DNA replication, DNA packaging, chromatin silencing and gene silencing pathways, opening potential novel targets for future precision medicine. [ABSTRACT FROM AUTHOR]

In this case report we describe a 30 year-old Caucasian woman with a histopathological diagnosis of pulmonary adenocarcinoma in both lungs with lipidic and micropapillary growth pattern and ROS1 (C-ros oncogene 1, receptor tyrosine kinase) rearrangement. There is evidence that crizotinib can be used for molecular target therapy in these patients. We enrolled the patient in an off-label program for the treatment of ROS1 rearranged adenocarcinomas with the EML4/anaplastic lymphoma kinase inhibitor crizotinib. After a follow-up of eight weeks we saw a complete remission in both lungs without any signs of metabolic tumor activity. This report shows the importance of testing young patients with adenocarcinomas of the lung for rare oncogenic driver mutations, such as ROS1, with possible molecular treatment options. [ABSTRACT FROM AUTHOR]

Recently the superiority of Crizotinib to standard first-line pemetrexed-plus-platinum chemotherapy in patients with previously untreated advanced ALK-positive NSCLC has been demonstrated. We report of a 36-year-old never smoker with advanced squamous cell carcinoma, presenting with a left lower lobe lesion, N3 nodal disease and multiple metastases (pleura, adrenal, muscle, bone). Despite squamous histology we decided on molecular testing. IHC for ALK was positive and confirmatory fluorescent in situ hybridization showed translocation of ALK. Although there is little evidence on ALK rearrangements in squamous NSCLC we decided on first line treatment with Crizotinib (250 mg twice daily). Eight and twelve weeks after treatment initiation a whole-body FDG fusion PET/CT scan showed dramatic tumor response with little remaining metabolic uptake in the left lobe and a single bone lesion. This evidence raises the question whether ALK testing should be done in never smokers with squamous NSCLC in the absence of oncogenic driver mutations. To our knowledge, this the first report of first line treatment of full squamous ALK-positive NSCLC with crizotinib. [ABSTRACT FROM AUTHOR]

Immune-checkpoint blockade in front-line or second-line treatment improves survival in advanced non-small cell lung cancer (aNSCLC) when compared with chemotherapy alone. However, easily applicable predictive parameters are necessary to guide immune-checkpoint inhibition in clinical practice. In this retrospective bi-centric analysis, we investigated the impact of baseline patient and tumor characteristics on clinical outcome in aNSCLC patients treated with programmed cell death protein 1(PD-1)/programmed cell death ligand 1 (PD-L1) inhibitors. Between May 2015 and January 2018, 142 unselected consecutive NSCLC patients received PD-1/PD-L1 inhibitors during the course of disease. In multivariate analysis, we identified the Eastern Cooperative Oncology Group (ECOG) performance status (ECOG > 1 versus ECOG ≤ 1, HR: 3.23, 95%CI: 1.58–6.60, P = 0.001), baseline absolute lymphocyte count (ALC; high: >0.93 × 109/L versus low: ≤ 0.93 × 109/L, HR: 0.38, 95%CI: 0.23–0.62, P < 0.001), prior or concomitant anti-vascular endothelial growth factor (VEGF) targeting therapy (yes versus no, HR: 2.18, 95%CI: 1.15–4.14, P = 0.017) and TNM stage (IV versus III, HR: 4.18, 95%CI: 1.01–17.36, P = 0.049) as the most relevant parameters for survival. Neither antibiotic exposure (antibiotic-positive versus antibiotic-negative, HR: 0.90, 95%CI: 0.56–1.45, P = 0.675), nor PD-L1 expression on tumor cells (≥1% versus <1%, HR: 0.68, 95%CI: 0.41–1.13, P = 0.140) was associated with survival. Baseline ECOG performance status and ALC were associated with survival in aNSCLC patients treated with PD-1/PD-L1 inhibitors and assessment of these parameters could be suitable in clinical practice. [ABSTRACT FROM AUTHOR]

Immune-checkpoint blockade in front-line or second-line treatment improves survival in advanced non-small cell lung cancer (aNSCLC) when compared with chemotherapy alone. However, easily applicable predictive parameters are necessary to guide immune-checkpoint inhibition in clinical practice. In this retrospective bi-centric analysis, we investigated the impact of baseline patient and tumor characteristics on clinical outcome in aNSCLC patients treated with programmed cell death protein 1(PD-1)/programmed cell death ligand 1 (PD-L1) inhibitors. Between May 2015 and January 2018, 142 unselected consecutive NSCLC patients received PD-1/PD-L1 inhibitors during the course of disease. In multivariate analysis, we identified the Eastern Cooperative Oncology Group (ECOG) performance status (ECOG > 1 versus ECOG ≤ 1, HR: 3.23, 95%CI: 1.58–6.60, P = 0.001), baseline absolute lymphocyte count (ALC; high: >0.93 × 109/L versus low: ≤ 0.93 × 109/L, HR: 0.38, 95%CI: 0.23–0.62, P < 0.001), prior or concomitant anti-vascular endothelial growth factor (VEGF) targeting therapy (yes versus no, HR: 2.18, 95%CI: 1.15–4.14, P = 0.017) and TNM stage (IV versus III, HR: 4.18, 95%CI: 1.01–17.36, P = 0.049) as the most relevant parameters for survival. Neither antibiotic exposure (antibiotic-positive versus antibiotic-negative, HR: 0.90, 95%CI: 0.56–1.45, P = 0.675), nor PD-L1 expression on tumor cells (≥1% versus <1%, HR: 0.68, 95%CI: 0.41–1.13, P = 0.140) was associated with survival. Baseline ECOG performance status and ALC were associated with survival in aNSCLC patients treated with PD-1/PD-L1 inhibitors and assessment of these parameters could be suitable in clinical practice. [ABSTRACT FROM AUTHOR]

Background: Induction chemotherapy incorporating docetaxel, cisplatin and 5- fluorouracil before radiotherapy may improve the outcome of patients with advanced head and neck cancer. Nevertheless, the addition of docetaxel increases hematological toxicity and infectious complications. Therefore, genetic markers predicting toxicity and efficacy of this treatment regimen may help to identify patients, who would have the most benefit from this intensive treatment.Methods: A cohort of 78 patients with advanced head and neck cancer treated with induction chemotherapy was assessed for clinical outcome and toxicity during treatment with curative intention. Genetic polymorphisms primary associated with treatment efficacy (ERCC2-rs13181, rs1799793, ERCC1-rs3212986, rs11615, XRCC1-rs25487) or with docetaxel caused toxicity (CYP39A1-rs7761731, SLCO1B3-rs11045585) were evaluated in all patients. The results of these analyses were correlated with the clinical outcome of the patients (loco regional control, progression free survival, overall survival) and treatment related toxicity during induction chemotherapy.Results: Median progression free survival and overall survival was 20 and 31 months in an intention to treat analysis, respectively. Overall response rate to induction chemotherapy was high with 78.1 % of all patients. None of the polymorphisms tested was associated with the clinical outcome of the patients. Genotype A of the CYP39A1 rs7761731 polymorphism was associated with a higher incidence of leucopenia and infections or death during induction chemotherapy.Conclusions: Intensive induction chemotherapy results in a high response rate in the majority of patients. None of the polymorphisms tested was associated with the clinical outcome of the patients. The CYP39A1 polymorphism rs7761731 may help to identify patients at high risk for treatment related toxicity. [ABSTRACT FROM AUTHOR]

Introduction: Restoration of function by dental implants following complete tumor removal has high priority in tumor surgery. Recurrent squamous cell carcinoma (SCC) after this procedure led us to search for possible causes. Method: 16 patients with primary oral SCC had undergone resection, postop. radiotherapy, various reconstructive procedures and functional rehabilitation by endosseous mandibular implants. In eight patients a recurrence was observed. The charts of these patients were examined for possible causal factors. Results: The primary SCC had always been located in the floor of the mouth and/or the mandibular alveolar process, and had been removed with sound margins. The time interval between primary resection and the placement of implants was 18–118months (mean 43mos). The recurrent SCC occurred in a similar area 1–69months (mean 34mos) after implantation. This area was again resected in seven patients 23–95months after the first resection (mean 65mos). Three patients died within 2–17months (mean 7mos) due to tumor progression. One patient died after 4months because of pulmonary embolism. The follow-up period of the other four patients (three tumor-free, one alive with disease) was 6–38months (mean 26mos). Conclusion: In our opinion the following factors may play a role: Close resection margins with remaining epithelial dysplasia, field cancerization, chronic inflammation due to poor oral hygiene, passive smoking, loss of sensitivity in the mandible after continuity resection, clinical misinterpretation of SCC as “granulation tissue”, HPV infection etc. The low number of patients does not allow unequivocal conclusions. We recommend a small mucosal biopsy at the time of implant placement. Symptoms of periimplantitis in former tumor patients should be considered as a serious sign. Short-term controls are mandatory as patients may not suffer from discomfort due to the loss of sensitivity. [Copyright &y& Elsevier]