Anti-angiogenic therapy using the multi-tyrosine kinase inhibitor Regorafenib enhances tumor progression in a transgenic mouse model of ß-cell carcinogenesis

Background: Pancreatic neuroendocrine tumors (PNETs) represent a distinct hypervascularized tumor entity, often diagnosed at metastatic stage. Therapeutic efficacy of anti-angiogenic multi-kinase inhibitors is frequently limited by primary or acquired resistance in vivo. This study aimed to characterize the molecular mode of action as well as resistance mechanisms to the anti-angiogenic multi-tyrosine kinase inhibitor (TKI) Regorafenib in vitro and in vivo. Methods: In vitro, human and murine pancreatic neuroendocrine cell lines were comparatively treated with Regorafenib and other TKIs clinically used in PNETs. Effects on cell viability and proliferation were analyzed. In vivo, transgenic RIP1Tag2 mice were treated with Regorafenib at two different time periods during carcinogenesis and its impact on angiogenesis and tumor progression was evaluated. Results: Compared to the established TKI therapies with Sunitinib and Everolimus, Regorafenib showed the strongest effects on cell viability and proliferation in vitro, but was unable to induce apoptosis. Unexpectedly and in contrast to these in vitro findings, Regorafenib enhanced proliferation during early tumor development in RIP1Tag2 mice and had no significant effect in late tumor progression. In addition, invasiveness was increased at both time points. Mechanistically, we could identify an upregulation of the pro-survival protein Bcl-2, the induction of the COX2-PGE2-pathway as well as the infiltration of CSF1R positive immune cells into the tumors as potential resistance mechanisms following Regorafenib treatment. Discussion: Our data identify important tumor cell-autonomous and stroma-dependent mechanisms of resistance to antiangiogenic therapies

Praxis und Herausforderungen der Delegation ärztlicher Tätigkeiten im interprofessionellen Arbeitsalltag der stationären Krankenversorgung in Deutschland: eine explorative Befragung

Hintergrund Die Delegation ärztlicher Tätigkeiten (DäT) hat im Alltag der stationären Behandlung eine hohe Bedeutung, stellt jedoch auch ein potenzielles Spannungsfeld der interprofessionellen Zusammenarbeit dar. Die Erfassung der Rahmenbedingungen der DäT im Arbeitsalltag ist Voraussetzung für die Beschreibung eines Optimierungspotenzials. Methode In einer bundesweiten explorativen Studie wurde im Bereich der stationären Versorgung tätiges ärztliches und nicht-ärztliches Personal mittels einer standardisierten Online-Befragung zu Aspekten der DäT befragt. Ergebnisse Von 757 Teilnehmer:innen (ärztlich 64,9 %) wurde DäT mehrheitlich als wirtschaftlich und zeitökonomisch sinnvoll (88,5 %) sowie im Interesse der Patient:innen (74 %) erachtet. Für 78,7 % der Befragten stellt DäT ein Spannungsfeld im Arbeitsalltag dar, abhängig von der Qualität der interprofessionellen Kommunikation. Eine unzureichende personelle Ausstattung wurde von 83,8 % als hinderlich für eine umfassendere DäT genannt. 63,2 % schätzten ihre Kenntnisse über rechtliche Aspekte der DäT als mindestens gut ein (Teilnehmer:innen mit < 5 Jahren Berufserfahrung: 52,6 %). Entsprechende Kenntnisse werden von ärztlichen Teilnehmer:innen überwiegend im Berufsalltag erworben (ärztlich 71,3 % vs. nicht-ärztlich 39,5 %). Schlussfolgerung DäT wurde über alle Berufsgruppen hinweg für sinnvoll erachtet, auch im Interesse der Patient:innen. Eine gezielte Förderung der sicheren und wirtschaftlichen DäT sollte bereits während der Ausbildung erfolgen. Eine höhere Ausschöpfung der Vorteile der DäT erfordert klare rechtliche Rahmenbedingungen, ein gut kommunizierendes Team und insbesondere eine angemessene personelle Ausstattung bei den Berufsgruppen, die delegierte Tätigkeiten übernehmen

Induction of oxidative- and endoplasmic-reticulum-stress dependent apoptosis in pancreatic cancer cell lines by DDOST knockdown

The dolichyl-diphosphooligosaccharide-protein glycosyltransferase non-catalytic subunit (DDOST) is a key component of the oligosaccharyltransferase complex catalyzing N-linked glycosylation in the endoplasmic reticulum lumen. DDOST is associated with several cancers and congenital disorders of glycosylation. However, its role in pancreatic cancer remains elusive, despite its enriched pancreatic expression. Using quantitative mass spectrometry, we identify 30 differentially expressed proteins and phosphopeptides (DEPs) after DDOST knockdown in the pancreatic ductal adenocarcinoma (PDAC) cell line PA-TU-8988T. We evaluated DDOST / DEP protein–protein interaction networks using STRING database, correlation of mRNA levels in pancreatic cancer TCGA data, and biological processes annotated to DEPs in Gene Ontology database. The inferred DDOST regulated phenotypes were experimentally verified in two PDAC cell lines, PA-TU-8988T and BXPC-3. We found decreased proliferation and cell viability after DDOST knockdown, whereas ER-stress, ROS-formation and apoptosis were increased. In conclusion, our results support an oncogenic role of DDOST in PDAC by intercepting cell stress events and thereby reducing apoptosis. As such, DDOST might be a potential biomarker and therapeutic target for PDAC

Translation, adaptation, and validation of the Care Coordination Instrument for cancer patients

Background: Cancer requires interdisciplinary intersectoral care. The Care Coordination Instrument (CCI) captures patients’ perspectives on cancer care coordination. We aimed to translate, adapt, and validate the CCI for Germany (CCI German version). Methods: The original English version contains 29 items in three domains, measured on a 4-point Likert scale (strongly disagree to strongly agree). Validation was conducted in three phases (mixed methods): (I) translation; (II) adaptation: pilot testing and revision in an iterative process using semi-structured, cognitive interviews with patients and professionals (physicians specializing in cancer), with interviews transcribed and qualitatively analyzed by inductive coding; and (III) validation: quantitative validation performed online (LimeSurvey), of at least 80 German patients, each with common cancer (breast, prostate) and rare cancer (different entities), with examination of factor structure (factor analysis) and determination of internal consistency (Cronbach's α) as well as potential influencing factors such as gender, education, or migration background (multivariable regression). Results: Six patients and six professionals tested the translated instrument for comprehensibility, readability, and acceptability. Two items were consistently problematic for interviewees. A 31-item version (29 items + 2 alternative items) was validated in 192 patients. The alternative items had a higher variance in response behavior and were better understood; therefore, they replaced the two problematic items. However, the three original domains could not be confirmed statistically. Exploratively, a two-factorial structure (with cross-loadings) emerged, which can be interpreted as “communication/information” (16 items) and “need-based navigation” (17 items). Overall, the instrument had a high internal consistency (total score α = 0.931, M = 47.16, SD = 14.25; communication/information α = 0.924, M = 30.14, SD = 8.93; need-based navigation α = 0.868, M = 23.99, SD = 8.37). Significant factors on the care coordination score are treatment location (hospital vs. private practice oncologist M = -9.83 score points, p = 0.011) and gender (women vs. men M = 8.92 score points, p = 0.002). Conclusion: The CCI German version is a valid instrument for measuring patients’ perceptions of cancer care coordination. Both domains reflect important aspects of care. The sensitivity of the CCI should be examined in future studies involving different cancer entities

Depletion of Macrophages Improves Therapeutic Response to Gemcitabine in Murine Pancreas Cancer.

BACKGROUND: The tumor microenvironment (TME) is composed of fibro-inflammatory cells and extracellular matrix (ECM) components. However, the exact contribution of the various TME compartments towards therapeutic response is unknown. Here, we aim to dissect the specific contribution of tumor-associated macrophages (TAMs) towards drug delivery and response in pancreatic ductal adenocarcinoma (PDAC). METHODS: The effect of gemcitabine was assessed in human and murine macrophages, human pancreatic stellate cells (hPSCs), and tumor cells (L3.6pl, BxPC3 and KPC) in vitro. The drug metabolism of gemcitabine was analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Preclinical studies were conducted using KrasG12D;p48-Cre and KrasG12D;p53172H;Pdx-Cre mice to investigate gemcitabine delivery at different stages of tumor progression and upon pharmacological TAM depletion. RESULTS: Gemcitabine accumulation was significantly increased in murine PDAC tissue compared to pancreatic intraepithelial neoplasia (PanIN) lesions and healthy control pancreas tissue. In vitro, macrophages accumulated and rapidly metabolized gemcitabine resulting in a significant drug scavenging effect for gemcitabine. Finally, pharmacological TAM depletion enhanced therapeutic response to gemcitabine in tumor-bearing KPC mice. CONCLUSION: Macrophages rapidly metabolize gemcitabine in vitro, and pharmacological depletion improves the therapeutic response to gemcitabine in vivo. Our study supports the notion that TAMs might be a promising therapeutic target in PDAC

Mental health in Germany in the first weeks of the Russo-Ukrainian war

Background: In the connected world, although societies are not directly involved in a military conflict, they are exposed to media reports of violence. Aims: We assessed the effects of such exposures on mental health in Germany during the military conflict in Ukraine. Method: We used the German population-based cohort for digital health research, DigiHero, launching a survey on the eighth day of the Russo-Ukrainian war. Of the 27 509 cohort participants from the general population, 19 444 (70.7%) responded within 17 days. We measured mental health and fear of the impact of war compared with other fears (natural disasters or health-related). Results: In a subsample of 4441 participants assessed twice, anxiety in the population (measured by the Generalised Anxiety Disorder-7 screener) was higher in the first weeks of war than during the strongest COVID-19 restrictions. Anxiety was elevated across the whole age spectrum, and the mean was above the cut-off for mild anxiety. Over 95% of participants expressed various degrees of fear of the impact of war, whereas the percentage for other investigated fears was 0.47–0.82. A one-point difference in the fear of the impact of war was associated with a 2.5 point (95% CI 2.42–2.58) increase in anxiety (11.9% of the maximum anxiety score). For emotional distress, the increase was 0.67 points (0.66–0.68) (16.75% of the maximum score). Conclusions: The population in Germany reacted to the Russo-Ukrainian war with substantial distress, exceeding reactions during the strongest restrictions in the COVID-19 pandemic. Fear of the impact of war was associated with worse mental health

Ketogenic diet as a glycine lowering therapy in nonketotic hyperglycinemia and impact on brain glycine levels

BACKGROUND: Nonketotic hyperglycinemia (NKH) is a severe neurometabolic disorder characterized by increased glycine levels. Current glycine reduction therapy uses high doses of sodium benzoate. The ketogenic diet (KD) may represent an alternative method of glycine reduction. AIM: We aimed to assess clinical and biochemical effects of two glycine reduction strategies: high dose benzoate versus KD with low dose benzoate. METHODS: Six infants with NKH were first treated with high dose benzoate therapy to achieve target plasma glycine levels, and then switched to KD with low dose benzoate. They were evaluated as clinically indicated by physical examination, electroencephalogram, plasma and cerebral spinal fluid amino acid levels. Brain glycine levels were monitored by magnetic resonance spectroscopy (MRS). RESULTS: Average plasma glycine levels were significantly lower with KD compared to benzoate monotherapy by on average 28%. Two infants underwent comparative assessments of brain glycine levels via serial MRS. A 30% reduction of brain glycine levels was observed in the basal ganglia and a 50% reduction in the white matter, which remained elevated above normal, and was equivalent between the KD and high dose benzoate therapies. CSF analysis obtained while participants remained on the KD showed a decrease in glycine, serine and threonine levels, reflecting their gluconeogenetic usage. Clinically, half the patients had seizure reduction on KD, otherwise the clinical impact was variable. CONCLUSION: KD is an effective glycine reduction method in NKH, and may provide a more consistent reduction in plasma glycine levels than high-dose benzoate therapy. Both high-dose benzoate therapy and KD equally reduced but did not normalize brain glycine levels even in the setting of low-normal plasma glycine