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Progression of prostate cancer (PCa) is characterized by metastasis and castration resistance after response to androgen deprivation. Therapeutic options are limited, causing high morbidity and lethality. Recent work reported pro-oncogenic implications of the Mediator subunits cyclin-dependent kinase (CDK) 8 and 19 for the progression of PCa. The current study explored the underlying molecular mechanisms of CDK8/CDK19 and tested effects of novel CDK8/CDK19 inhibitors. PC3, DU145, LNCaP, and androgen-independent LNCaP Abl were used for in vitro experiments. Two inhibitors and CDK19 overexpression were used to modify CDK8/CDK19 activity. MTT assay, propidium iodide staining, wound healing assay, Boyden chamber assay, and adhesion assay were used to investigate cell viability, cell cycle, migration, and adhesion, respectively. Peptide-kinase screen using the PamGene platform was conducted to identify phosphorylated targets. Combining CDK8/CDK19 inhibitors with anti-androgens led to synergistic antiproliferative effects and sensitized androgen-independent cells to bicalutamide. CDK8/CDK19 inhibition resulted in reduced migration and increased collagen I-dependent adhesion. Phosphorylation of multiple peptides linked to cancer progression was identified to be dependent on CDK8/CDK19. In summary, this study substantially supports recent findings on CDK8/CDK19 in PCa progression. These findings contribute to a better understanding of underlying pro-oncogenic effects, which is needed to develop CDK8/CDK19 as a therapeutic target in PCa.

To verify the feasibility of the SF-MPF for oral reconstruction, the anatomic, sonographic and histologic features of the SF-MPF were investigated and the outcome in a series of patients was evaluated. The sonographic and histologic results showed a supra-fascial arterio-venous vascular blood supply to the sub-fascial design of the MPF. The clinical course of 12 consecutive patients who underwent oral reconstruction using the SF-MPF along with ipsi- or lateral neck dissection for treatment of oral cancer showed sufficient pedicle length and reliable blood supply. The SF-MPF is a reliable and safe pedicled myocutaneous flap. Therefore, it should be considered being an additional option when a pedicled flap has to be selected.

MAPK signaling pathways are constitutively active in colon cancer and also promote acquired resistance to MEK1 inhibition. Here, we demonstrate that BRAFV600E-mutated colorectal cancers acquire resistance to MEK1 inhibition by inducing expression of the scaffold protein CEMIP through a β-catenin– and FRA-1–dependent pathway. CEMIP was found in endosomes and bound MEK1 to sustain ERK1/2 activation in MEK1 inhibitor–resistant BRAFV600E-mutated colorectal cancers. The CEMIP-dependent pathway maintained c-Myc protein levels through ERK1/2 and provided metabolic advantage in resistant cells, potentially by sustaining amino acids synthesis. CEMIP silencing circumvented resistance to MEK1 inhibition, partly, through a decrease of both ERK1/2 signaling and c-Myc. Together, our data identify a cross-talk between Wnt and MAPK signaling cascades, which involves CEMIP. Activation of this pathway promotes survival by potentially regulating levels of specific amino acids via a Myc-associated cascade. Targeting this node may provide a promising avenue for treatment of colon cancers that have acquired resistance to targeted therapies.Significance: MEK1 inhibitor–resistant colorectal cancer relies on the scaffold and endosomal protein CEMIP to maintain ERK1/2 signaling and Myc-driven transcription. Cancer Res; 78(16); 4533–48. ©2018 AACR.

Figure S1. Selumetinib-resistant organoids show elevated mRNA levels of CEMIP but not Myc. Figure S2. CEMIP controls the expression of genes candidates linked to ErbB, RAS and LEF1 signaling and identification of upstream regulators of CEMIP. Figure S3. CEMIP is required for the maintenance of ex-vivo organoid cultures showing constitutive Wnt signaling. Figure S4. CEMIP promotes the acquired resistance to MEK1 inhibition in colon cancer cells. Figure S5. Ectopic expression of CEMIP in colon cancer cells enhances pERK1/2 and Myc levels and protects from cell death upon MEK1 inhibition. Figure S6. Acquisition of Selumetinib resistance in BRAFV600E-mutated colorectal cancer cells decreases E-cadherin levels and induces the nuclear expression of p65 and FRA-1. Figure S7. FRA-1 controls CEMIP expression in Selumetinib-resistant colon cancer cells. Figure S8. Vemurafenib decreases CEMIP expression in Selumetinib-resistant colon cancer cells. Figure S9. CEMIP expression is regulated by TCF4 in Selumetinib-resistant colon cancer cells. Figure S10. Generation of KRASG13D or G12A-mutated colon cancer cells showing some acquired resistance to Selumetinib. Figure S11. CEMIP promotes the acquired resistance to Selumetinib in BRAFV600E-mutated colorectal cancer cells. Figure S12. pERK1/2 levels decrease after a short treatment with Selumetinib but increase again after 24 hours of MEK1 inhibition and CEMIP contributes to MEK1-dependent ERK1/2 activation. Figure S13. Resistant BRAFV600E-mutated colorectal cancer cells with acquired resistance to MEK1 inhibition show more BRAF dimers. Figure S14. Myc deficiency mimicks CEMIP deficiency in Selumetinib-resistant ex-vivo organoids. Supplementary Table 1 is a list of all antibodies used in this study.

Legends to Supplementary Figures S1-S8.

1.5 x 105 HeLa cells/6-well were either left untransfected or were transfected with (A) 0 µg, 0.025 µg or 0.1 µg of pCMV-Flag2-IRF1 or (B) 0 µg, 0.6 µg or 0.8 µg of pCMV-Flag2-IRF2.

Neoadjuvant radio/chemotherapy regimens can markedly improve cervical cancer outcome in a subset of patients, while other patients show poor responses, but may encounter severe adverse effects. Thus, there is a strong need for predictive biomarkers to improve clinical management of cervical cancer patients. STAT3 is considered as a critical antiapoptotic factor in various malignancies. We therefore investigated STAT3 activation during cervical carcinogenesis and its impact on the response of cervical cancer cells to chemotherapeutic drugs. Tyr705-phosphorylated STAT3 increased from low-grade cervical intraepithelial neoplasia (CIN1) to precancerous CIN3 lesions. Notably, pTyr705-STAT3 activation significantly declined from CIN3 to invasive cancer, also when compared in the same clinical biopsy. pTyr705-STAT3 was also low or absent in cultured human cervical cancer cell lines, consistent with the in vivo expression data. Unexpectedly, IL6-type cytokine signaling inducing STAT3 activation rendered cervical cancer cells significantly more susceptible to chemotherapeutic drugs, that is, cisplatin or etoposide. This chemosensitization was STAT3-dependent and we identified IFN regulatory factor-1 (IRF1) as the STAT3-inducible mediator required for cell death enhancement. In line with these data, pTyr705-STAT3 significantly correlated with nuclear IRF1 expression in cervical cancer in vivo. Importantly, high IRF1 expression in pretreatment cervical cancer biopsy cells was associated with a significantly better response to neoadjuvant radio/chemotherapy of the patients. In summary, our study has identified a key role of the STAT3/IRF1 pathway for chemosensitization in cervical cancer. Our results suggest that pretherapeutic IRF1 expression should be evaluated as a novel predictive biomarker for neoadjuvant radio/chemotherapy responses. Cancer Res; 76(13); 3872–83. ©2016 AACR.

The recent characterization of a group of non-MYC rearranged aggressive B-cell lymphomas, resembling Burkitt lymphoma (BL), characteristically harboring a telomeric 11q loss or combined 11q proximal gains/loss pattern has led to the introduction of the provisional entity of Burkitt-like lymphoma with 11q aberration (BLL-11q). Prompted by the discovery of a telomeric 11q loss in an HIV+ high-grade B-cell lymphoma patient, we investigated an extended cohort of aggressive B-cell lymphomas, enriched for cases with histopathological features intermediate between DLBCL and BL, including double- and triple-hit lymphomas (n = 47), for 11q loss/combined 11q proximal gains/loss pattern by fluorescence in situ hybridization. We provide first evidence that 11q aberrations can be found in both BLL in the context of an underlying HIV infection as well as in high-grade B-cell lymphomas with MYC, BCL2, and/or BCL6 rearrangements. We therefore propose that the clinicopathological spectrum of malignancies carrying this aberration may be broader than previously assumed.

Background/aim We investigated the prevalence of human papillomavirus (HPV) in a prospective cohort of patients with squamous cell carcinoma of the oral cavity (OSCC) using both p16INK4a and HPV DNA, i.e., double positivity, as a definition criterion. Additionally, we examined the association of HPV with survival. Patients and methods Samples from 280 OSCC patients were analyzed for HPV-positivity using p16INK4a immunohistochemistry (IHC) and in situ hybridization (ISH)/LCD arrays, for HPV low and high-risk types. Only patients positive for both p16INK4a and HPV DNA were considered as HPV-positive. Survival probabilities and 95% confidence intervals were estimated using the Kaplan-Meier method. Cox proportional hazards models were used to assess HPV association with disease-free survival (DFS), cause-specific survival (CSS) and overall survival (OS) in a competing risks scenario. Results Specimen from 30 (10.7%) patients were p16+ and HPV DNA+, while 31 (11.0%) were either p16+ or HPV DNA+ only. OS probabilities at five years for HPV-positive and -negative groups were 50.9% (35.4%-73.1%) and 52.9% (47.0%-59.5%), respectively. HPV double positivity influenced neither OS, CSS nor DFS: HR=0.84 (0.43-1.63), 1.64 (0.76-3.54) and 1.13 (0.55-2.35), respectively. Conclusion In contrast to oropharyngeal cancer, the prevalence of HPV in OSCC is low and the presence of HPV does not influence survival outcomes. Hence, there is no evidence to support a parallel transfer of therapy regimen for HPV-positive OPC to OSCC, in terms of therapy de-escalation and/or vaccination.

Surgery remains the only curative treatment of pancreatic neuroendocrine neoplasms (pNEN). Here, we report the outcome after surgery for non-functional pNEN at a European Neuroendocrine Tumor Society (ENETS) center in Germany between 2000 and 2019, cases were analyzed for surgical (Clavien–Dindo classification, CDc) and oncological outcomes. Forty-nine patients (tumor grading G1 n = 25, G2 n = 22, G3 n = 2), with a median age of 56 years, were included. Severe complications (CDc ≥ grade 3b) occurred in 11 patients (22.4%) and type B/C pancreatic fistulas (POPFs) occurred in 5 patients (10.2%), in-hospital mortality was 2% (n = 1). Six of seven patients with tumor recurrence (14.3%) had G2 tumors in the pancreatic body/tail. The median survival was 5.7 years (68 months, [1–228 months]). Neither the occurrence (p = 0.683) nor the severity of complications had an influence on the relapse behavior (p = 0.086). This also applied for a POPF (≥B, p = 0.609). G2 pNEN patients (n = 22) with and without tumor recurrence had similar median tumor sizes (4 cm and 3.9 cm, respectively). Five of the six relapsed G2 patients (83.3%) had tumor-positive lymph nodes (N+), all G2 pNEN patients with recurrence had initially been treated with distal pancreatic resection. Pancreatic resections for pNEN are safe but associated with relevant postoperative morbidity. Future studies are needed to evaluate suitable resection strategies for G2 pNEN.

ZusammenfassungDie Instillationstherapie stellt im Feld der urothelialen Neoplasien der Harnblase eine einzigartige Therapiemöglichkeit dar, die so in keinem anderen medizinischen Bereich zur Anwendung kommt. Mit der Wahl des zu instillierenden Medikamentes hat sich die Forschung der vergangenen Jahre intensiv beschäftigt, hierbei hat sich besonders der Einsatz von Bacillus Calmette-Guerin (BCG) und Mitomycin C (MMC) bewährt. Weitere Medikamente, die jedoch seltener zum Einsatz kommen, sind Chemotherapeutika wie Gemcitabin, dessen klinische Relevanz in diesem Artikel beleuchtet werden soll.So vielfältig, wie die Therapiemöglichkeiten der Harnblasentumore, ist auch ihre Histopathologie. Das invertierte Papillom stellt im Formenkreis der urologischen Erkrankungen eine seltene Krankheitsentität dar. Eine Assoziation zum nicht-muskelinvasiven Harnblasenkarzinom wird diskutiert.Wir berichten von dem Fall einer Patientin mit invertiertem Papillom der Harnblase, welche nach Versagen einer BCG-Therapie und einer allergischen Reaktion gegen MMC mit einer Gemcitabin-Instillationstherapie behandelt wurde.

A 54-year-old patient with a history of pulmonary tuberculosis and occupational exposure to dust in early adulthood presented with symptoms of coughing with sputum, weight loss, occasional night sweats, and thoracic pain. Non-necrotizing granulomatosis in lung and lymph-node biopsies indicated sarcoidosis. Combined immunosuppressive therapy did not lead to an improvement. An atypical lung resectate with fibroinflammatory changes and obliterative endothelialitis may finally lead to the diagnosis of IgG4-associated lung disease with a bronchovascular pattern of involvement. The question discussed here is whether this is a coexistence of IgG4-associated lung disease with sarcoidosis or the spectrum of one disease.

Background In salivary-gland-carcinomas (SGC), there is only a small fraction of entities that appears to profit from immune-checkpoint-inhibition (ICI). Recent findings connected the activation of adenosine-signaling with a tolerogenic microenvironment. Therefore, the inhibition of adenosine-pathway markers (CD39 and/or CD73) can augment ICI and/or display a novel immunotherapeutic strategy beyond ICI. Here, we assessed the immunohistochemical expression of CD39 and CD73 across a wide spectrum of SGCs. Methods In total, 114 patients with SGCs consecutively diagnosed between 2001 and 2021 were assessed for clinicopathological baseline-characteristics and underwent confirmatory histopathological review. Immunohistochemical expression-levels of CD39 and CD73 were assessed by applying the tumor-proportion-score (TPS) and the immune-proportional-score (IPS) comparable to PD-L1 expression analysis in routine clinical practice. Additionally, findings were correlated with PD-L1 expression-levels. Results The median age was 60.6 and 51.8% patients were female. The cohort covered a spectrum of eight distinct entities. Advanced-stage disease (UICC/AJCC III/IVA-IVC) at initial diagnosis was present in the majority of patients (64/114). Immunohistochemical-staining revealed positivity for CD39 and CD73 in 48.2% and 21.1% on tumor-cells (TPS ≥ 1%) as well as 46.4% and 42.9% within the immune-cell infiltrate (IPS ≥ 1%), respectively. Further comparative analyses revealed immune-cold entities such adenoidcystic-carcinoma (AdCC), immune-hot tumors such as adenocarcinoma, not otherwise specified (AC (NOS)) and entities with intermediate immunologic features such as acinic-cell-carcinoma (ACC). Conclusion Current results indicate entity-specific adenosine-signaling signatures. These findings suggest that the adenosine-pathway plays a decisive role in tumor immunity among the major spectrum of SGCs. Targeting the adenosine-pathway might pose a promising therapeutic option for selected entities.

Die Informationen aus pathologischen Befundberichten zu Prostatastanzbiopsien stellen im Management des Prostatakarzinoms einen essentiellen Bestandteil fur die Risikostratifizierung dar. Durch zusatzliche Parameter ist der Informationsgehalt der Befunde zunehmend komplexer geworden. Mit Hilfe einer reprasentativen Umfrage zu pathologischen Befundberichten von Prostatastanzbiopsien sollte ermittelt werden, wie diese von Urologen interpretiert und fur Therapieentscheidungen berucksichtigt werden. Uber den Berufsverband der Deutschen Urologen e. V. (BDU) wurde ein Fragebogen zu Prostatastanzbiopsiebefunden an seine niedergelassenen Mitglieder verschickt. Bei Abschluss der Studie lagen 109 Antworten, davon 106 auswertbare Fragebogen, vor. Die Anzahl positiver Stanzen und der prozentuale Tumoranteil werden hinsichtlich der Tumorausdehnung fur die Risikostratifizierung als relevanteste Angaben angesehen. Bei mp(multiparametrischen)MRT-Fusionsstanzen wird eine separate Angabe zur Anzahl positiver Stanzen innerhalb des Standardschemas und der mpMRT-positiven Foci bevorzugt. Am haufigsten wird die Stanze mit dem hochsten Gleason-Score berucksichtigt, wobei dem prozentualen Gleason-4-Muster auch eine wichtige Bedeutung zukommt. Etwa die Halfte der Befragten wunscht sich auch die Angabe der neuen „grade groups“ nach ISUP/WHO (International Society of Urological Pathology/World Health Organization). Als weitere gewunschte Parameter werden u. a. der Ki67-Proliferationsindex, der PSMA(„prostate-specific membrane antigen“)-Status sowie Angaben zu intraduktalen oder neuroendokrinen Tumorkomponenten aufgefuhrt. Fur die Angaben histopathologischer Parameter im Befundbericht von Prostatastanzbiopsien existiert kein einheitlicher Standard und bei der Interpretation gibt es eine Variabilitat. Weitere Studien und eine klare Aussage seitens der Leitlinien werden notig sein, um ein einheitliches Diagnose‑/Prognoseschema zu etablieren.

We compared postoperative radiotherapy (PORT) to surgery only (SO), and supraomohyoidal neck dissection (SOHND) to modified radical neck dissection (MRND) in patients with pT1-T2 squamous cell carcinomas of the oral cavity (OSCC) and a single cervical lymph node metastasis (pN1) in terms of overall survival (OS), oral cancer specific survival (OCSS), and regional recurrence-free survival (RRFS), in a prospective cohort study.We included patients with pT1-T2 pN1 OSCC with no distant metastasis and estimated the survival probabilities using the Kaplan-Meier method and calculated hazards ratios (HR) for PORT vs. SO and MRND vs. SOHND using adjusted Cox regression models.A total of 51 patients (26 SO vs. 25 PORT, 9 SOHND vs. 42 MRND) were evaluated. Patients who received PORT were more likely to be younger and healthier. OS at 5 years was 41% and 87% in the SO and PORT groups, respectively. OS at 5 years was 52% and 67% in the in the SOHND and MRND groups, respectively. Both OCSS and RRFS were improved by PORT. Extending neck dissection was not associated with improved OS (HR = 0.83).PORT is associated with preferable OS, OCSS, and RRFS in pT1-2 pN1 oral cancer and should be recommended regularly.

We investigated the prevalence of human papillomavirus (HPV) in a prospective cohort of patients with squamous cell carcinoma of the oral cavity (OSCC) using both p16Samples from 280 OSCC patients were analyzed for HPV-positivity using p16Specimen from 30 (10.7%) patients were p16+ and HPV DNA+, while 31 (11.0%) were either p16+ or HPV DNA+ only. OS probabilities at five years for HPV-positive and -negative groups were 50.9% (35.4%-73.1%) and 52.9% (47.0%-59.5%), respectively. HPV double positivity influenced neither OS, CSS nor DFS: HR=0.84 (0.43-1.63), 1.64 (0.76-3.54) and 1.13 (0.55-2.35), respectively.In contrast to oropharyngeal cancer, the prevalence of HPV in OSCC is low and the presence of HPV does not influence survival outcomes. Hence, there is no evidence to support a parallel transfer of therapy regimen for HPV-positive OPC to OSCC, in terms of therapy de-escalation and/or vaccination.

BACKGROUND Guidelines recommended for resection of oral cancer define a free margin of ≥5 mm as clear and safe (R0). This statement was questioned recently based on the assumption that different surgical margins may hold different risk categories. The aim of this study was to investigate the impact of stratification of the surgical margins on the survival outcome of patients with oral cancer. METHODS In a cohort of 753 patients, the hazard ratio for local recurrence-free survival (LRFS), overall survival (OS), and oral cancer-specific survival (OCSS) were estimated for R0 resection, the close margin of 1-4 mm, involved resection borders but with free frozen sections. Competing risk factors were considered in the statistical regression model. RESULTS One hundred seventy-three (23%) patients developed local recurrence and 316 (42%) died in the 5 follow-up years. There was a gradual improvement in the LRFS, OCSS, OS with the increase of clear margin. OS showed a similar tendency. CONCLUSION Not all patients with an R0cm status carry the same risk for impaired LRFS, OCSS, and OS. Their risk to develop recurrence is higher than those patients with R0 ≥5 mm but stratified risk management can be recommended according to the presented results.

Summary High-grade prostatic intraepithelial neoplasia (HGPIN) is a facultative precursor lesion of prostate cancer (PCa). Multifocal HGPIN in needle biopsies in the absence of PCa indicates a higher risk of cancer detection in subsequent biopsies. Therefore, a reliable diagnosis of HGPIN is of high clinical relevance guiding the management of patients with cancer-negative biopsies. Detection of HGPIN is merely based on morphological features while biomarkers aiding in the diagnosis of HGPIN and its differentiation from benign glands and other glandular lesions are lacking yet. Here, we investigated the expression of cyclin-dependent kinase 19 (CDK19) by immunohistochemistry on prostate needle biopsies of 140 patients who were all diagnosed with PCa using whole-tissue sections and compared CDK19 levels between HGPIN, PCa, and adjacent benign glands. In addition, CDK19 was compared with AMACR expression in a subset of intraductal carcinomas (IDCs) on radical prostatectomy (RP) specimens. HGPIN was present in 65.7% of biopsies and in 88% associated to adjacent PCa. CDK19 overexpression defined as moderate to high CDK19 expression visible at low magnification was found in 82.6% of HGPIN. In contrast, 89.3% of benign glands were CDK19-negative or demonstrated only low CDK19 expression highlighting a high sensitivity and specificity to accurately detect HGPIN based on CDK19 expression levels. CDK19 was overexpressed in 59% of PCa but did not correlate significantly with the expression of intermingled HGPIN. On RP, CDK19 and AMACR showed no significant difference in the detection rate of IDC. In summary, assessment of CDK19 facilitates accurate and simplified diagnosis of HGPIN with high sensitivity and specificity and aides the differentiation to non-neoplastic glandular alterations. Considering the high clinical significance of diagnosis HGPIN that still has a limited reproducibility among pathologists, we suggest CDK19 as diagnostic biomarker for HGPIN.

Neck tumors are rare fetal conditions. They can receive their growth stimuli by activating missense mutations characterizing disorders with benign overgrowth, collectively known as PIK3CA-Related-Overgrowth-Spectrum. This results in segmental overgrowth with phenotypic variation, genetic heterogeneity or tissue specific distribution. Thus, clinical and molecular diagnosis as well as treatment remains challenging.

Intravesical chemotherapy instillation is a unique method of treatment confined to urothelial neoplasia. Within the last decades, the substances Bacillus Calmette-Guerin (BCG) and mitomycin C (MMC) have evolved as the standard regimens for intravesical therapy. However, there are other chemotherapeutic substances, which are used less frequently, such as gemcitabine. In this article we aim to highlight the clinical relevance of intravesical gemcitabine instillations as treatment of non-muscle invasive bladder cancer.The histological subtypes of bladder tumours are as diverse as the intravesical regimens. Inverted papilloma is a rare entity in the spectrum of urological diseases. There seems to be an association with non-muscle invasive bladder cancer.We report a rare case of an inverted bladder papilloma treated with intravesical gemcitabine instillations after BCG failure and an allergic reaction to MMC.Die Instillationstherapie stellt im Feld der urothelialen Neoplasien der Harnblase eine einzigartige Therapiemöglichkeit dar, die so in keinem anderen medizinischen Bereich zur Anwendung kommt. Mit der Wahl des zu instillierenden Medikamentes hat sich die Forschung der vergangenen Jahre intensiv beschäftigt, hierbei hat sich besonders der Einsatz von Bacillus Calmette-Guerin (BCG) und Mitomycin C (MMC) bewährt. Weitere Medikamente, die jedoch seltener zum Einsatz kommen, sind Chemotherapeutika wie Gemcitabin, dessen klinische Relevanz in diesem Artikel beleuchtet werden soll.So vielfältig, wie die Therapiemöglichkeiten der Harnblasentumore, ist auch ihre Histopathologie. Das invertierte Papillom stellt im Formenkreis der urologischen Erkrankungen eine seltene Krankheitsentität dar. Eine Assoziation zum nicht-muskelinvasiven Harnblasenkarzinom wird diskutiert.Wir berichten von dem Fall einer Patientin mit invertiertem Papillom der Harnblase, welche nach Versagen einer BCG-Therapie und einer allergischen Reaktion gegen MMC mit einer Gemcitabin-Instillationstherapie behandelt wurde.

Purpose: Recurrent tumors (RT) of head and neck squamous cell carcinoma (HNSCC) occur in up to 60%, with poor therapeutic response and detrimental prognosis. We hypothesized that HNSCC RTs successfully evade antitumor immune response and aimed to reveal tumor immune microenvironment (TIME) changes of primary tumors (PT) and corresponding RTs. Experimental Design: Tumor-infiltrating leukocytes (TIL) of 300 PTs and 108 RTs from two large independent and clinically well-characterized HNSCC cohorts [discovery cohort (DC), validation cohort (VD)] were compared by IHC. mRNA expression analysis of 730 immune-related genes was performed for 18 PTs and RTs after adjuvant chemoradiotherapy (CRT). The effect of chemotherapy and radiation resistance was assessed with an in vitro spheroid/immunocyte coculture model. Results: TIME analysis revealed overall decrease of TILs with significant loss of CD8+ T cells (DC P = 0.045/VC P < 0.0001) and B lymphocytes (DC P = 0.036/VC P < 0.0001) in RTs compared with PTs in both cohorts. Decrease predominantly occurred in RTs after CRT. Gene expression analysis confirmed loss of TILs (P = 0.0004) and B lymphocytes (P < 0.0001) and showed relative increase of neutrophils (P = 0.018), macrophages (P < 0.0001), dendritic cells (P = 0.0002), and mast cells (P = 0.0057) as well as lower overall expression of immune-related genes (P = 0.018) in RTs after CRT. Genes involved in B-lymphocyte functions and number of tertiary lymphoid structures showed the strongest decrease. SPP1 and MAPK1 were upregulated in vivo and in vitro, indicating their potential suitability as therapeutic targets in CRT resistance. Conclusions: HNSCC RTs have an immunosuppressive TIME, which is particularly apparent after adjuvant CRT and might substantially contribute to poor therapeutic response and prognosis.

A 54-year-old patient with a history of pulmonary tuberculosis and occupational exposure to dust in early adulthood presented with symptoms of coughing with sputum, weight loss, occasional night sweats, and thoracic pain. Non-necrotizing granulomatosis in lung and lymph-node biopsies indicated sarcoidosis. Combined immunosuppressive therapy did not lead to an improvement. An atypical lung resectate with fibroinflammatory changes and obliterative endothelialitis may finally lead to the diagnosis of IgG4-associated lung disease with a bronchovascular pattern of involvement. The question discussed here is whether this is a coexistence of IgG4-associated lung disease with sarcoidosis or the spectrum of one disease.Ein 54 Jahre alter Patient mit anamnestisch im frühen Erwachsenenalter nicht austherapierter Lungentuberkulose und beruflicher Staubexposition wird mit dem Beschwerdebild von Husten mit Auswurf, Gewichtsverlust, gelegentlichem Nachtschweiß und thorakalen Schmerzen vorstellig. Eine nichtnekrotisierende Granulomatose in Lungen- und Lymphknotenbiopsaten weist auf eine Sarkoidose hin. Unter kombinierter immunsuppressiver Therapie kommt es zu keiner Befundbesserung. Anhand eines atypischen Lungenresektates mit fibroinflammatorischen Veränderungen und obliterativer Endothelialitis kann schlussendlich eine IgG4-assoziierte Lungenerkrankung vom bronchovaskulären Typ diagnostiziert werden. Besprochen wird hier die Frage, ob es sich um eine Koexistenz einer IgG4-assoziierten Lungenerkrankung mit einer Sarkoidose handelt oder um das Spektrum einer Erkrankung.

Summary: Cdkn1a, which encodes p21, functions as a major route for p53-mediated cell-cycle arrest. However, the consequence of Cdkn1a gene dosage on tumor suppression has not been systematically investigated. Here, we employed BAC transgenesis to generate a Cdkn1aSUPER mouse, which harbors an additional Cdkn1a allele within its natural genomic context. We show that these mice display enhanced cell-cycle arrest and reduced apoptosis in response to genotoxic stress. Furthermore, using a chemically induced skin cancer model and an autochthonous Kras-driven lung adenocarcinoma model, we show that Cdkn1aSUPER mice display a cancer protection phenotype that is indistinguishable from that observed in Tp53SUPER animals. Moreover, we demonstrate that Tp53 and Cdkn1a cooperate in mediating cancer resistance, using a chemically induced fibrosarcoma model. Overall, our Cdkn1aSUPER allele enabled us to assess the contribution of Cdkn1a to Tp53-mediated tumor suppression. : Torgovnick et al. create a mouse model, carrying a third copy of Cdkn1a (p21), which shows enhanced cell-cycle arrest capacity and protection against DNA damage-induced apoptosis. The Cdkn1aSUPER animals display delayed epithelial regeneration and a robust cancer resistance phenotype, highlighting the importance of p21 in p53-dependent tumor suppression. Keywords: Cdkn1a, p21, p53, mouse model, cancer, tumor suppressor, cell cycle arrest, apoptosis, cancer protection

FGFR1 amplification has been found in 15% of patients with breast cancer and has been postulated as a promising marker to predict response against FGFR inhibitors. However, early phase clinical trials of selective FGFR inhibitors demonstrated only limited efficacy in FGFR1-amplified breast cancer patients. We found that BGJ398, an FGFR inhibitor, effectively inhibited phosphorylation of FGFR1 and MEK/ERK signaling in FGFR1-amplified breast cancer without affecting tumor cell proliferation. However, FGFR1 knockout inhibited tumor angiogenesis in vivo. We unraveled that FGFR1 regulates the secretion of the proangiogenic vascular endothelial growth factor (VEGF) in a MAPK-dependent manner. We further found that FGF-FGFR1 signaling induces an autocrine activation of VEGF-VEGFR1 pathway that again amplifies VEGF secretion via VEGF-VEGFR1-AKT signaling. Targeting both VEGFR1 and FGFR1 resulted in synergistic anti-angiogenic treatment effects in vivo. We thus postulate synergistic treatment effects in FGFR1/VEGFR1-positive breast cancer patients by dual targeting of FGFR and VEGFR.

// Heike Loeser 1, *, ** , Simon Schallenberg 1, *, ** , Moritz von Winterfeld 1 , Lars Tharun 1 , Hakan Alakus 3, * , Arnulf Holscher 4 , Elfriede Bollschweiler 3 , Reinhard Buettner 1 , Thomas Zander 2, *, ** and Alexander Quaas 1, *, ** 1 Institute of Pathology, University of Cologne, Cologne, Germany 2 Department I of Internal Medicine, Center for Integrated Oncology (CIO), University of Cologne, Cologne, Germany 3 Department of General, Visceral and Cancer Surgery, University of Cologne, Cologne, Germany 4 Department of Thorax and Oesophageal Surgery, Agaplesion Markus Krankenhaus, Frankfurt/Main, Germany * Gastrointestinal Cancer Group Cologne (GCGC) ** These authors have contributed equally to this work Correspondence to: Alexander Quaas, email: alexander.quaas@uk-koeln.de Keywords: esophageal adenocarcinoma; TUBB3; RNA-in-situ-hybridization; immunohistochemistry Received: May 10, 2017 Accepted: November 20, 2017 Published: December 11, 2017 ABSTRACT Background: Esophageal adenocarcinomas show an increasing incidence in the Western world and their overall survival remains low. Microtubules are multifunctional cytoskeletal proteins involved in crucial cellular roles, including maintenance of cell shape, intracellular transport, meiosis, and mitosis. Microtubulus-TUBB3 was found overexpressed in several carcinomas suggesting a significant role in cancer development. High levels of TUBB3 expression were also described to be associated with poor clinical outcome in various cancers. It was shown that overexpression of TUBB3 could be related to reduced efficiency of taxane-based targeting anticancer drugs in several cancer types. Results: There is a statistically significant association between high TUBB3 protein and TUBB3 mRNA expression and shortened survival (p

// Axel Lechner 1, 2, * , Hans Schloser 1, 3, * , Sacha I. Rothschild 1, 4, * , Martin Thelen 1 , Sabrina Reuter 1 , Peter Zentis 5 , Alexander Shimabukuro-Vornhagen 1, 6 , Sebastian Theurich 1, 6, 7 , Kerstin Wennhold 1 , Maria Garcia-Marquez 1 , Lars Tharun 8 , Alexander Quaas 8 , Astrid Schauss 5 , Jorg Isensee 9 , Tim Hucho 9 , Christian Huebbers 10 , Michael von Bergwelt-Baildon 1, 6, * and Dirk Beutner 2, * 1 Cologne Interventional Immunology, Department I of Internal Medicine, University Hospital of Cologne, Cologne, Germany 2 Department of Otorhinolaryngology, Head and Neck Surgery, University of Cologne, Cologne, Germany 3 Department of General, Visceral and Cancer Surgery, University of Cologne, Cologne, Germany 4 University Hospital Basel, Department of Internal Medicine, Medical Oncology, Basel, Switzerland 5 Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), Cologne, Germany 6 Department I of Internal Medicine, Center for Integrated Oncology (CIO), University Hospital of Cologne, Cologne, Germany 7 Max-Planck-Institute for Metabolism Research, Cologne, Germany 8 Institute of Pathology, University of Cologne, Cologne, Germany 9 Department of Anesthesiology and Intensive Care Medicine, Experimental Anesthesiology and Pain Research, University Hospital of Cologne, University of Cologne, Germany 10 Jean-Uhrmacher Institute for Clinical ENT Research, University of Cologne, Cologne, Germany * These authors contributed equally to this work Correspondence to: Michael von Bergwelt-Baildon, email: michael.bergwelt@uk-koeln.de Keywords: squamous cell carcinoma, head and neck, microenvironment, T cells, immune checkpoint Received: January 10, 2017 Accepted: April 25, 2017 Published: May 16, 2017 ABSTRACT The composition of tumor-infiltrating lymphocytes (TIL) reflects biology and immunogenicity of cancer. Here, we characterize T-cell subsets and expression of immune checkpoint molecules in head and neck squamous cell carcinoma (HNSCC). We analyzed TIL subsets in primary tumors ( n = 34), blood (peripheral blood mononuclear cells (PBMC); n = 34) and non-cancerous mucosa ( n = 7) of 34 treatment-naive HNSCC patients and PBMC of 15 healthy controls. Flow cytometry analyses revealed a highly variable T-cell infiltration mainly of an effector memory phenotype (CD45RA − /CCR7 − ). Naive T cells (CD45RA + /CCR7 + ) were decreased in the microenvironment compared to PBMC of patients, while regulatory T cells (CD4 + /CD25 + /CD127 low and CD4 + /CD39 + ) were elevated. Furthermore, we performed digital image analyses of entire cross sections of HNSCC to define the ‘Immunoscore’ (CD3 + and CD8 + cell infiltration in tumor core and invasive margin) and quantified MHC class I expression on tumor cells by immunohistochemistry. Immune checkpoint molecules cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4), programmed cell death 1 (PD-1) and programmed cell death 1 ligand 1 (PD-L1) were increased in TILs compared to peripheral T cells in flow-cytometric analysis. Human papillomavirus (HPV) positive tumors showed higher numbers of TILs, but a similar composition of T-cell subsets and checkpoint molecule expression compared to HPV negative tumors. Taken together, the tumor microenvironment of HNSCC is characterized by a strong infiltration of regulatory T cells and high checkpoint molecule expression on T-cell subsets. In view of increasingly used immunotherapies, a detailed knowledge of TILs and checkpoint molecule expression on TILs is of high translational relevance.

Introduction Multiparametric magnetic resonance imaging (mpMRI)/ultrasound fusion-guided biopsy, in short “targeted biopsy (TB)”, is becoming more attractive as it improves the detection of clinically significant prostate cancer (CaP). The accuracy of fusion-guided biopsies is limited due to false positive radiological findings as well as to histological evidence for cancer in radiologically inconspicuous regions of the prostate. We aimed to analyze histomorphological findings on mpMRI lesions highly suspicious for CaP classified as PI-RADS 4 or PI-RADS 5 (Prostate Imaging - Recording and Data System) but cancer-negative in the biopsy of this region of interest (ROI), and to compare them with findings in radiologically inconspicuous regions. Materials and methods We re-evaluated prostate biopsies from 57 patients who underwent TB in combination with systematic standard biopsy (SB) from June 2017 to July 2018 at the University Hospital Schleswig Holstein Campus Luebeck. Out of 143 ROIs, 34 PI-RADS 4/5 cancer-negative lesions were identified and subjected to comprehensive histomorphological reevaluation. Contralateral cancer-negative SBs were used as control. Chi-square test was used for statistical analysis. Results The frequency of histomorphological alterations including stromal, glandular, vascular, and inflammatory alterations were 97% and 79.2% in prostatic tissues from cancer-negative TBs and SBs, respectively. Stromal, glandular, and inflammatory alterations were present in the majority of biopsies from both TBs and SBs. Statistical analysis revealed no significant difference between TBs and SBs with regard to stromal, glandular, and inflammatory alterations. However, vascular abnormalities were exclusively detected in TBs (18.2%). Conclusion The frequency of histomorphological alterations is slightly higher in prostate tissues from TBs compared to SB. Only vascular alterations seem to be distinct for TBs. However, it has to be assumed that additional factors influence the false-negative rate of mpMRI/ultrasound fusion-guided TB.

Delaunay et al. reveal the role of U34 tRNA-modifying enzymes in the regulation of specific mRNA translation to support cell invasion and metastasis., Quantitative and qualitative changes in mRNA translation occur in tumor cells and support cancer progression and metastasis. Posttranscriptional modifications of transfer RNAs (tRNAs) at the wobble uridine 34 (U34) base are highly conserved and contribute to translation fidelity. Here, we show that ELP3 and CTU1/2, partner enzymes in U34 mcm5s2-tRNA modification, are up-regulated in human breast cancers and sustain metastasis. Elp3 genetic ablation strongly impaired invasion and metastasis formation in the PyMT model of invasive breast cancer. Mechanistically, ELP3 and CTU1/2 support cellular invasion through the translation of the oncoprotein DEK. As a result, DEK promotes the IRES-dependent translation of the proinvasive transcription factor LEF1. Consistently, a DEK mutant, whose codon composition is independent of U34 mcm5s2-tRNA modification, escapes the ELP3- and CTU1-dependent regulation and restores the IRES-dependent LEF1 expression. Our results demonstrate that the key role of U34 tRNA modification is to support specific translation during breast cancer progression and highlight a functional link between tRNA modification– and IRES-dependent translation during tumor cell invasion and metastasis.

Neoadjuvant radio/chemotherapy regimens can markedly improve cervical cancer outcome in a subset of patients, while other patients show poor responses, but may encounter severe adverse effects. Thus, there is a strong need for predictive biomarkers to improve clinical management of cervical cancer patients. STAT3 is considered as a critical antiapoptotic factor in various malignancies. We therefore investigated STAT3 activation during cervical carcinogenesis and its impact on the response of cervical cancer cells to chemotherapeutic drugs. Tyr705-phosphorylated STAT3 increased from low-grade cervical intraepithelial neoplasia (CIN1) to precancerous CIN3 lesions. Notably, pTyr705-STAT3 activation significantly declined from CIN3 to invasive cancer, also when compared in the same clinical biopsy. pTyr705-STAT3 was also low or absent in cultured human cervical cancer cell lines, consistent with the in vivo expression data. Unexpectedly, IL6-type cytokine signaling inducing STAT3 activation rendered cervical cancer cells significantly more susceptible to chemotherapeutic drugs, that is, cisplatin or etoposide. This chemosensitization was STAT3-dependent and we identified IFN regulatory factor-1 (IRF1) as the STAT3-inducible mediator required for cell death enhancement. In line with these data, pTyr705-STAT3 significantly correlated with nuclear IRF1 expression in cervical cancer in vivo. Importantly, high IRF1 expression in pretreatment cervical cancer biopsy cells was associated with a significantly better response to neoadjuvant radio/chemotherapy of the patients. In summary, our study has identified a key role of the STAT3/IRF1 pathway for chemosensitization in cervical cancer. Our results suggest that pretherapeutic IRF1 expression should be evaluated as a novel predictive biomarker for neoadjuvant radio/chemotherapy responses. Cancer Res; 76(13); 3872–83. ©2016 AACR.

Hintergrund Neben der klassischen Diagnosestellung ist der Pathologe zunehmend mit der Tumorsubtypisierung und Ermittlung therapierelevanter Biomarker beim Magenkarzinom beauftragt.

Dickkopf-2 (DKK2) has been described as Wnt/beta-catenin pathway antagonist and its expression is mediated by micro RNA-221 (miRNA-221). So far, there is only limited data characterizing the role of DKK2 expression in esophageal cancer. A tissue micro array of 192 patients with esophageal adenocarcinoma was analyzed immunohistochemically for DKK2, miRNA-221 expression by RNA scope, and GATA6 amplification by fluorescence in-situ hybridization. The data was correlated with clinical, pathological and molecular data (TP53, HER2, c-myc, GATA6, PIK3CA, and KRAS amplifications). DKK2 expression was detectable in 21.7% and miRNA-221 expression in 33.5% of the patients. We observed no correlation between DKK2 or miRNA-221 expression and clinico-pathological data DKK2 expression was correlated with TP53 mutations and amplification of GATA6. We did not detect a survival difference in dependence of DKK2 for the total cohort, however, in patients without neoadjuvant treatment DKK2 expression correlated with a prolonged survival (median overall-survival 202 vs. 55 months, p = 0.012) which turned opposite in patients that underwent neoadjuvant treatment. High amounts of miRNA-221 were in trend associated with a prolonged overall-survival (p = 0.070). DKK2 as a Wnt antagonist is associated with prolonged survival in patients without neoadjuvant treatment and changes its prognostic value to the contrary in patients after neoadjuvant therapy. The modulatory effects of neoadjuvant treatment in connection with DKK2 expression are not fully understood, but when considering DKK2 as a tumor marker, it is necessary to see it in the context of neoadjuvant therapy.

Reprogramming of mRNA translation has a key role in cancer development and drug resistance(1). However, the molecular mechanisms that are involved in this process remain poorly understood. Wobble tRNA modifications are required for specific codon decoding during translation(2,3). Here we show, in humans, that the enzymes that catalyse modifications of wobble uridine 34 (U-34) tRNA (U-34 enzymes) are key players of the protein synthesis rewiring that is induced by the transformation driven by the BRAF(V600E) oncogene and by resistance to targeted therapy in melanoma. We show that BRAF(V600E)-expressing melanoma cells are dependent on U34 enzymes for survival, and that concurrent inhibition of MAPK signalling and ELP3 or CTU1 and/or CTU2 synergizes to kill melanoma cells. Activation of the PI3K signalling pathway, one of the most common mechanisms of acquired resistance to MAPK therapeutic agents, markedly increases the expression of U-34 enzymes. Mechanistically, U-34 enzymes promote glycolysis in melanoma cells through the direct, codon-dependent, regulation of the translation of HIF1A mRNA and the maintenance of high levels of HIF1 alpha protein. Therefore, the acquired resistance to anti-BRAF therapy is associated with high levels of U-34 enzymes and HIF1 alpha. Together, these results demonstrate that U-34 enzymes promote the survival and resistance to therapy of melanoma cells by regulating specific mRNA translation.