147 research outputs found
Entschlüsselung der metastatischen Evolution von Urothelkarzinomen - Proteom- und Transkriptom-Profilierung zur Ermittlung von Therapieresistenzmechanismen
No full textHintergrund und Ziele: Die metastatische Evolution des Urothelkarzinoms, die zu
klonaler Heterogenität durch den Erwerb verschiedener neuer (Treiber-)Veränderungen
bzw. Veränderungen der Tumormikroumgebung führt ist ein aktueller
Forschungsschwerpunktes des Feldes. Zahlreiche bisherige Arbeiten indizieren eine
erhebliche molekulare Heterogenität und Evolution zwischen Primärtumoren und
Fernmetastasen, die einerseits intrinsisch aber auch extrinsisch (z.B. Therapiedruck)
angetrieben wird. In aktuellen klinischen und translationalen Studien wird der
metastatischen Evolution häufig allerdings wenig Beachtung geschenkt, und in der Regel
werden für die Bestimmung „präzisiononkologischer“ Biomarker innerhalb dieser Studien
Primärtumorproben verwendet, die den aktuellen molekularen und genetischen
Krankheitsstatus nur inadäquat widerspiegeln. Diese Verzerrung erklärt möglicherweise
auch die eingeschränkte Wertigkeit von Biomarkern, die in anderen Tumorentitäten
bereits standardmäßig für die Therapieprädiktion eingesetzt werden (z.B. molekulare
Subtypen beim Brustkrebs, PD-L1 bei Lungenkrebs). Ziel der vorliegenden Arbeit ist
daher gewesen, besser zu verstehen, wie sich bestimmte Tumoreigenschaften während
der Metastasierung verändern, insbesondere das immunologische Microenvironment,
molekluare Subypten und die Expression von wichtigen Oberflächenmolekülen wie
NECTIN-4; und wie die Veränderungen dieser biologischen Faktoren das Ansprechen
auf Immuntherapien, Chemotherapien und neue smarte Chemotherapien (Antikörper-
Drug-Konjugate) beeinflussen.
Patienten und Methoden: Im Rahmen der vorliegenden Arbeit wurden metastatische
Gewebeproben von 154 Patienten mit metastasiertem Urothelkarzinom (bei 138
Patienten mit verfügbaren gematchten Primärtumoren und Fernmetastasen) untersucht.
Mittels verschiedener Untersuchungstechnologien (Immunhistochemie, räumliche
Proteomik, RNAseq) wurden die gematchten Gewebeproben auf Veränderungen ihrer
immunologischen Mikroumgebungen (inkl. wichtiger Immuncheckpointproteine wie PDL1),
ihrer molekularen Subtypen und ihres NECTIN-4
Oberflächenproteinexpressionsstatus untersucht. Zusätzlich wurde eine Kohorte von
n=47 Patienten auf den NECTIN-4 Oberflächenexpressionsstatus untersucht, die mit
dem gegen NECTIN-4 gerichteten Antikörper-Drug-Konjugat Enfortumab Vedotin
behandelt wurden. Die Analyseresultate wurden mit Ansprechraten und dem Outcome
der Patienten unter Chemotherapie, Immuntherapie und unter Enfortumab Vedotin
korreliert.
Ergebnisse und Beobachtungen: Das immunologische Microenvironment inklusive der
Expression von PD-L1 auf Tumor- und Immunzellen unterscheidet sich deutlich zwischen
gematchten Primärtumoren und Fernmetastasen. Während der immunologische
Primärtumorstatus für das Ansprechen auf moderne Immuntherapien und
Chemotherapien weder prädiktiv noch prognostisch ist, ist der immunologische Status
der Metastase für das Ansprechen und für Langzeiterfolge unter Chemotherapie und
Immuntherapie entscheidend. Korrespondierend dazu konnten wir beobachten, dass
basale und luminale molekulare Tumorzellsubtypen/Lininedifferenzierungen während der
metastatischen Evolution stabil bleiben, aber die Tumore ihre Mikroumgebung (z.B. das
tumorassoziierte Bindegewebe oder das immunologische Microenvironment) in
Fernmetastasen stark remodellieren. Ebenso unterliegt auch der Expressionsstatus des
Oberflächenmarkers NECTIN-4 (Zelladhäsionsmolekül) einer relevanten Dynamik
während der metastatischen Evolution (Reduktion), und ist, wenn er an Metastasen
bestimmt wird, im Falle einer starken membranären Expression positiv prädiktiv für gute
Ansprechraten und lange Ansprechdauern auf Enfortumab Vedotin.
Schlussfolgerungen und Diskussion: Biologische Kerneigenschaften wie die
immunologische Mikroumgebung und die Oberflächenexpression von Angriffspunkten für
moderne Antikörper-Drugkonjugate wie NECTIN-4 verändern sich während der
metastatischen Evolution substantiell, während molekulare Kerneigenschaften wie die
molekulare Liniendifferenzierung (molekulare Tumorzellsubtypen) relativ stabil bleiben.
Die hoch dynamische Veränderung immunologischer Eigenschaften und der Expression
von NECTIN-4 in Fernmetastasen korreliert zudem stark mit dem Ansprechen auf
Immuntherapien und Enfortumab Vedotin. Wir schlussfolgern daher, dass die
Bestimmung präzisionsonkologischer Biomarker, insbesondere, wenn es sich dabei um
Marker der Zell-Zell-Interaktion handelt, in Fernmetastasengewebe bestimmt werden
sollte, und nicht in wenig repräsentativem Primärtumormaterial. Unsere vorliegenden
Ergebnisse müssen allerdings in Zukunft prospektiv validiert werden.Background and Objectives: The metastatic evolution of urothelial carcinoma, leading
to clonal heterogeneity through the acquisition of various new (driver) alterations or
changes in the tumor microenvironment, is a current research focus of the field.
Numerous previous studies indicated a significant molecular heterogeneity and evolution
between primary tumors and distant metastases, driven both intrinsically and extrinsically
(e.g., by therapy pressure). In current clinical and translational studies, however, the
metastatic evolution is often given little attention, and primary tumor samples are typically
used to determine "precision oncology" biomarkers, what inadequately reflects the current
molecular and genetic disease status. This may also explain the limited value of
biomarkers that are already routinely used for therapy prediction in other tumor entities
(e.g., molecular subtypes in breast cancer, PD-L1 in lung cancer). Therefore, the aim of
this study was to better understand how certain tumor characteristics change during
metastasis, especially the immunological microenvironment, molecular subtypes, and the
expression of important surface molecules like NECTIN-4; and how changes in these
biological factors influence the response to immunotherapies, chemotherapies, and new
smart chemotherapies (antibody-drug conjugates).
Patients and Methods: In the context of this study, metastatic tissue samples from 154
patients with metastatic urothelial carcinoma (with 138 patients having available matched
primary tumors and distant metastases) were examined. Using various examination
technologies (immunohistochemistry, spatial proteomics, RNAseq), the matched tissue
samples were examined for changes in their immunological environments (including
important immune checkpoint proteins like PD-L1), their molecular subtypes, and their
NECTIN-4 surface protein expression status. Additionally, a cohort of n=47 patients was
examined for the NECTIN-4 surface expression status, who were treated with the anti-
NECTIN-4 antibody-drug conjugate Enfortumab Vedotin. The analysis results were
correlated with response rates and patient outcomes under chemotherapy,
immunotherapy, and Enfortumab Vedotin.
Results and Observations: The immunological microenvironment, including the
expression of PD-L1 on tumor and immune cells, differs significantly between matched primary tumors and distant metastases. While the immunological status of the primary
tumor is neither predictive nor prognostic for the response to modern immunotherapies
and chemotherapies, the immunological status of the metastasis is crucial for the
response and long-term success under chemotherapy and immunotherapy.
Correspondingly, we observed that basal and luminal molecular tumor cell subtypes/line
differentiations remain stable during metastatic evolution, but the tumors strongly remodel
their microenvironment (e.g., the tumor-associated connective tissue or the
immunological microenvironment) in distant metastases. Similarly, the expression status
of the surface marker NECTIN-4 (cell adhesion molecule) undergoes relevant dynamics
during the metastatic evolution (reduction), and, when determined on metastases, is
positively predictive of good response rates and long durations of response to Enfortumab
Vedotin in case of strong membranous expression.
Conclusions and Discussion: Biological core properties such as the immunological
microenvironment and the surface expression of targets for modern antibody-drug
conjugates like NECTIN-4 change substantially during the metastatic evolution, while
molecular core properties such as molecular lineage differentiation (molecular tumor cell
subtypes) remain relatively stable. The highly dynamic change in immunological
properties and the expression of NECTIN-4 in distant metastases correlates strongly with
the response to immunotherapies and Enfortumab Vedotin. Therefore, we conclude that
the determination of precision oncological biomarkers, especially when it comes to
markers of cell-cell interaction, should be made in distant metastasis tissue, and not in
less representative primary tumor material. However, our current results must be
prospectively validated in the future
An integrated cellular and molecular model of gastric neuroendocrine cancer evolution highlights therapeutic targets
No full textSomatostatin receptor expression related to TP53 and RB1 alterations in pancreatic and extrapancreatic neuroendocrine neoplasms with a Ki67-index above 20⁒
No full textSomatostatin receptor 2A expression is a feature of well-differentiated neuroendocrine neoplasms and is important for their diagnosis and therapy. Little is known about somatostatin receptor 2A expression in poorly differentiated neuroendocrine neoplasms in relation to TP53 and RB1 status and how these features may contribute to the separation of well from poorly differentiated neuroendocrine neoplasms with a proliferation index above 20%. This study investigates the expression of somatostatin receptors, p53 and Rb1, and TP53 alterations in pancreatic and extrapancreatic well and poorly differentiated neuroendocrine neoplasms (Ki67-index >20%). Thirty-seven poorly differentiated neuroendocrine neoplasms of pancreatic (n=12) and extrapancreatic origin (n=25) as well as 10 well-differentiated neuroendocrine neoplasms of the pancreas (n=9) and rectum (n=1) with a Ki67-index >20% were immunostained for synaptophysin, chromogranin A, Ki67, CD56, p53, Rb1, ATRX, DAXX, progesterone receptor, somatostatin receptor 2A, somatostatin receptor 5, and cytokeratin 20, and sequenced for TP53, exons 5-9. Somatostatin receptor 2A was positive in 6/37 of poorly differentiated and in 8/10 of well-differentiated neuroendocrine neoplasms. One well-differentiated and two poorly differentiated neuroendocrine neoplasms expressed somatostatin receptor 5. Abnormal nuclear p53 and Rb1 staining was found in 29/37 and 22/37 poorly differentiated neuroendocrine neoplasms, respectively, whereas all well-differentiated neuroendocrine neoplasms showed normal p53 and Rb1 expression. TP53 gene alterations were restricted to poorly differentiated neuroendocrine neoplasms (24/34) and correlated well with p53 expression. All cases were progesterone receptor negative. Somatostatin receptor 2A expression is not limited to well-differentiated neuroendocrine neoplasms but also occurs in 16% of poorly differentiated neuroendocrine neoplasms from various sites. Most poorly differentiated neuroendocrine neoplasms are characterized by TP53 alterations and Rb1 loss, usually in the absence of somatostatin receptor 2A expression. In the pancreas, these criteria contribute to separate well-differentiated neuroendocrine neoplasms with a Ki67-index above 20% from poorly differentiated neuroendocrine neoplasms.Modern Pathology advance online publication, 6 January 2017; doi:10.1038/modpathol.2016.217
Pancreatic neuroendocrine carcinomas reveal a closer relationship to ductal adenocarcinomas than to neuroendocrine tumors G3
No full textPancreatic neuroendocrine carcinoma is a rare aggressive tumor commonly harboring TP53 and RB1 alterations and lacking neuroendocrine related genetic changes such as mutations in MEN1 and ATRX/DAXX. Little is known about its genetic profile with regard to that of pancreatic ductal adenocarcinoma. We therefore conducted a detailed genetic study in 12 pancreatic neuroendocrine carcinomas of large cell (n=9) and small cell type (n=3) using massive parallel sequencing applying a 409 gene panel on an Ion Torrent system. The genetic data were compared with known data of pancreatic ductal adenocarcinoma and correlated with exocrine lineage marker expression. A similar analysis was performed in 11 pancreatic neuroendocrine tumors G3. Neuroendocrine carcinomas harbored 63 somatic mutations in 45 different genes, affecting most commonly TP53 (8/12 cases), KRAS (5/12 cases) and RB1 (loss of expression with or without deletion in 4/12 cases). Five carcinomas had both TP53 and KRAS mutations. Neuroendocrine tumors G3 only shared singular mutations in five different genes with neuroendocrine carcinomas, including TP53, CDKN2A, ARID1A, LRP1B and APC, affecting five different cases. Most KRAS positive neuroendocrine carcinomas also expressed MUC1 (4/5) and carcinoembryonic antigen (3/5) as markers of ductal differentiation. Our data indicate that almost half of the pancreatic neuroendocrine carcinomas are genetically and phenotypically related to pancreatic ductal adenocarcinoma, and might therefore respond to chemotherapies targeting the latter carcinomas
Non-canonical functions of SNAIL drive context-specific cancer progression
No full textAbstract SNAIL is a key transcriptional regulator in embryonic development and cancer. Its effects in physiology and disease are believed to be linked to its role as a master regulator of epithelial-to-mesenchymal transition (EMT). Here, we report EMT-independent oncogenic SNAIL functions in cancer. Using genetic models, we systematically interrogated SNAIL effects in various oncogenic backgrounds and tissue types. SNAIL-related phenotypes displayed remarkable tissue- and genetic context-dependencies, ranging from protective effects as observed in KRAS- or WNT-driven intestinal cancers, to dramatic acceleration of tumorigenesis, as shown in KRAS-induced pancreatic cancer. Unexpectedly, SNAIL-driven oncogenesis was not associated with E-cadherin downregulation or induction of an overt EMT program. Instead, we show that SNAIL induces bypass of senescence and cell cycle progression through p16 INK4A -independent inactivation of the Retinoblastoma (RB)-restriction checkpoint. Collectively, our work identifies non-canonical EMT-independent functions of SNAIL and unravel its complex context-dependent role in cancer
DLL3 Expression in Neuroendocrine Carcinomas and Neuroendocrine Tumours: Insights From a Multicentric Cohort of 1294 Pulmonary and Extrapulmonary Neuroendocrine Neoplasms
No full textAbstract Delta-like ligand 3 (DLL3) is frequently expressed in pulmonary small cell neuroendocrine carcinoma (SCNEC) and has emerged as a promising therapeutic target. However, limited data on DLL3 expression in other neuroendocrine neoplasms (NEN), such as extrapulmonary SCNEC, large cell neuroendocrine carcinomas (LCNEC), mixed neuroendocrine-non-neuroendocrine neoplasms (MiNEN), gastroenteropancreatic neuroendocrine tumours (GEP-NET), and pulmonary carcinoids, impedes an estimation if other types of NEN might be suitable candidates for anti-DLL3 therapies. We evaluated DLL3 expression in 1294 NEN and 479 non-neuroendocrine carcinomas, correlating the findings with histological subtypes, tumour localisation, and overall survival (OS). Furthermore, we explored the concordance of DLL3 expression during metastatic progression in 67 paired primary NEN and metastases. DLL3 expression was significantly higher in NEC (64.0%) compared to GEP-NET and pulmonary carcinoids (10.1%, p < 0.001), particularly in SCNEC (80.4%), followed by LCNEC (62.6%) and MiNEN (28.6%). DLL3 was common in pulmonary carcinoids (41.5%), but rare in GEP-NET (5.1%) and non-neuroendocrine carcinomas (1.3%). Overall DLL3 expression was highly concordant between metastases and corresponding primary NEN (92.5%, p < 0.001). In univariable analyses, DLL3-expressing pulmonary carcinoids ( p = 0.005) and GEP-NET ( p = 0.018) were associated with decreased OS, but this was not retained in multivariable analyses adjusting for stage and grade ( p = n. s.). No prognostic impact was observed in pulmonary ( p = 0.708) or GEP-NEC ( p = 0.87). Our study highlights significant differences in DLL3 expression across NEN subtypes and localisations, with largely concordant expression in metastases. DLL3-based therapies may be effective in many NEC and pulmonary carcinoids, while DLL3 appears to be a minor therapeutic target for GEP-NET and non-neuroendocrine carcinomas.Bundesministerium für Bildung und Forschung https://doi.org/10.13039/501100002347Deutsche Krebshilfe https://doi.org/10.13039/501100005972Philipps-Universität Marburg 10000896
Genetische Analyse der Hämoxygenase-1 bei verschiedenen Formen der Pankreatitis
No full textDie Hämoxygenase-1 (HO-1) ist das geschwindigkeitsbestimmende Enzym des Hämabbaus und ist wichtiger Regulator inflammatorischer Prozesse. Der Verlauf einer experimentellen akuten Pankreatitis (AP) konnte im Tiermodell durch HO-1 Induktion abgemildert werden. Die Aktivierung und Proliferation pankreatischer Stellatum Zellen (PSC) wird durch eine experimentelle HO-1 Induktion inhibiert und kann so möglicherweise vor der Fibrosierung des Pankreasparenchyms bei chronischer Pankreatitis (CP) schützen. Die Transkription der HO-1 wird durch einen GT-Repeat beeinflusst, der im Promoter lokalisiert ist. Diese Arbeit untersuchte, ob Varianten des GT-Repeat oder weitere genetische Varianten der HO-1 mit verschiedenen Pankreatitisformen assoziiert sind.
Der GT-Repeat und der SNP rs2071746 wurden mit fluoreszensmarkierten Primern bzw. mit Schmelzkurvenanalyse bei 285 Patienten mit AP, bei 208 Patienten mit alkoholischer CP (ACP), bei 207 mit idiopathischer/hereditärer CP (ICP/HCP), 147 Patienten mit Alkoholischer Leberzirrhose (ALZ) und bei 289 Kontrollen untersucht. Bei den ACP Patienten wurde die GT-Repeat Analyse auf insgesamt 446 Patienten erhöht. Zusätzlich wurden die kodierenden HO-1 Abschnitte mittels DNA-Sequenzierung bei 145 Patienten mit ACP, 138 Patienten mit ICP/HCP, 147 Patienten mit ALZ und bei 151 Kontrollen analysiert. Das Exon 3 wurde darüber hinaus bei zusätzlichen ICP/HP Patienten und Kontrollen untersucht.
Die Längenverteilungen des GT-Repeat, die Allelverteilung des SNP rs2071746 und die Verteilung der bei der DNA-Sequenzierung gefundenen synonymen und nicht synonymen Varianten waren bei allen untersuchten Gruppen nicht signifikant unterschiedlich.
Obwohl die funktionellen Daten einen Einfluss von HO-1 Varianten auf die Pathogenese der verschiedenen Pankreatitis-Formen nahelegen, konnte unsere umfangreiche genetische Analyse keine Assoziation nachweisen. Genetische Varianten der HO-1 haben keinen Einfluss auf die Entwicklung einer AP, ACP, ICP/HCP und ALZ.:Inhaltsverzeichnis
Vorbemerkung ..................................................................................................... 3
Bibliographische Beschreibung.......................................................................... 4
Abkürzungen/Abbildungen ................................................................................ 6
1. Einleitung........................................................................................................9 1.1 Akute Pankreatitis ......................................................................................................................... 9
1.2 Chronische Pankreatitis ............................................................................................................... 11 1.3 Genetische Aspekte der Chronischen Pankreatitis ...................................................................... 12 1.3.1 Kationisches Trypsinogen (PRSS1) ...................................................................................... 12 1.3.2 Anionisches Trypsinogen (PRSS2) ....................................................................................... 14 1.3.3 Serinproteaseinhibitor, Kazal Typ1 (SPINK1)..................................................................... 14 1.3.4 Chymotrypsin C (CTRC) ...................................................................................................... 15 1.3.5 CFTR (Cystic Fibrosis Transmembrane Conductance Regulator) ...................................... 15 1.4 Hämoxygenase-1 ......................................................................................................................... 16 1.4.1 Physiologische Bedeutung der Hämoxygenase-1 (HO-1) .................................................... 16 1.4.2 Genetische Varianten der Hämoxygenase-1 ........................................................................ 18 1.4.3 Hämoxygenase-1 und Pankreatitis....................................................................................... 20 1.5 Hypothese/Fragestellung ............................................................................................................. 21
2. Publikation ..................................................................................................... 22 3. Zusammenfassung der Arbeit ...................................................................... 23 4. Literaturverzeichnis...................................................................................... 28 5. Danksagung.................................................................................................... 35 6. Erklärung über die eigenständige Abfassung der Arbeit .......................... 36 7. Lebenslauf ...................................................................................................... 3
Monitoring conflict risk: The contribution of globally used indicator systems
No full textThe Chapter deals with the contribution of globally used indicator systems, in particular governance, environmental and MDG indicators, to the measurement of the risk of violent conflict. The author argues that instead of specialising on conflict indicators in a narrow sense, one should rather go for a highly public and transparent broad indicator set along the lines of a Sustainable Development Index; the reason being that only a mainstream index can exert media pressure on political actors.JRC.DG.G.3 - Econometrics and applied statistic
On the art of aggregating "apples & oranges"
No full textThe Communication on Environmental Indicators and Green National Accounting (Com (94) 670 final, 21.12.94) laid the basis for indicator and accounting projects aimed to give comprehensive support to environmental policy, like GARP and GreenStamp. The article is intended to demonstrate: that indicators are a powerful driving force of many, if not most, political decisions; that bad indicators are thus a recipe for bad politics; how an indicator system that serves democratic decision-making should be designed. In particular, in order to avoid serious policy distortions, environmental policy needs a broad spectrum of indicators covering all relevant issues. Valuation projects that treat pollutants with sophisticated methodologies like Impact Pathway Analysis (GARP) or Avoidance Cost-based economic modelling (GreenStamp) face data availability problems which force them in practice to concentrate on selected pollutants, excluding all others; the result is policy distortion. The author proposes that valuation methodologies should focus on general patterns, like the dependence of Avoidance Costs from implementation speed, type of instruments, and level of analysis (i.e. company, sector, national economy, EU economy), instead of treating single pollutants with unnecessary precision while neglecting many others. Policy Performance Indices (PPI) may serve as an instrument to extend such general valuations to a broad range of pollutants and policy questions, using numéraires plus relative weighting as the basis for valuations
Neuroendocrine neoplasms of the pancreas: diagnosis and pitfalls
No full textCommon to neuroendocrine neoplasms of the pancreas is their expression of synaptophysin, chromogranin A, and/or INSM1. They differ, however, in their histological differentiation and molecular profile. Three groups can be distinguished: well-differentiated neuroendocrine neoplasms (neuroendocrine tumors), poorly differentiated neuroendocrine neoplasms (neuroendocrine carcinomas), and mixed neuroendocrine-non-neuroendocrine neoplasms. However, the expression of synaptophysin and, to a lesser extent, also chromogranin A is not restricted to the neuroendocrine neoplasms, but may also be in a subset of non-neuroendocrine epithelial and non-epithelial neoplasms. This review provides the essential criteria for the diagnosis of pancreatic neuroendocrine neoplasms including diagnostic clues for the distinction of high-grade neuroendocrine tumors from neuroendocrine carcinomas and an algorithm avoiding diagnostic pitfalls in the delineation of non-neuroendocrine neoplasms with neuroendocrine features from pancreatic neuroendocrine neoplasms
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