Competing sorting signals guide endolyn along a novel route to lysosomes in MDCK cells

We have examined the trafficking of the mucin-like protein endolyn in transfected, polarized MDCK cells using biochemical approaches and immunofluorescence microscopy. Although endolyn contains a lysosomal targeting motif of the type YXXΦ and was localized primarily to lysosomes at steady state, significant amounts of newly synthesized endolyn were delivered to the apical cell surface. Antibodies to endolyn, but not lamp-2, were preferentially internalized from the apical plasma membrane and efficiently transported to lysosomes. Analysis of endolyn–CD8 chimeras showed that the lumenal domain of endolyn contains apical targeting information that predominates over basolateral information in its cytoplasmic tail. Interestingly, surface polarity of endolyn was independent of O-glycosylation processing, but was reversed by disruption of N-glycosylation using tunicamycin. At all times, endolyn was soluble in cold Triton X-100, suggesting that apical sorting was independent of sphingolipid rafts. Our data indicate that a strong, N-glycan-dependent apical targeting signal in the lumenal domain directs endolyn into a novel biosynthetic pathway to lysosomes, which occurs via the apical surface of polarized epithelial cells

Cancer rehabilitation support by cancer counselling centres (CARES): study protocol of a quasi-experimental feasibility study

Introduction While maintaining or restoring work ability after a cancer diagnosis is an essential aim of the rehabilitation process for working-age patients, problems can arise during the return to work (RTW) or when retaining work. Counselling could provide support for patients with or after cancer with employment-related questions (eg, questions related to RTW and work retention). Outpatient psychosocial cancer counselling centres in Germany offer counselling on work-related questions; however, resources for this are limited. This protocol presents a feasibility study of an intensified needs-based counselling intervention that supports those seeking employment-related advice.Methods and analysis The CARES (cancer rehabilitation support by cancer counselling centres) project is a feasibility study for a newly developed counselling intervention. The intervention is being developed as part of the project and piloted in about 20 outpatient cancer counselling centres. The CARES study has a quasi-experimental pre–post design with a control cohort. First, patients who undergo regular counselling are recruited. Second, after the counsellors have been trained for the newly developed intervention, participants for the intervention group are recruited from the cancer counselling centres. Quantitative and formative evaluations will be performed in accordance with the existing guidelines. The quantitative evaluation comprises three patient surveys (at the beginning of the counselling process, 3 months into the counselling process and, for the intervention group, at the end of the counselling process) and routine data of the counselling process. The formative evaluation includes interviews with patients, counsellors and other stakeholders, as well as participatory observations of counselling sessions.Ethics and dissemination Approval has been obtained from the ethics committee of the Medical Faculty of the University Bonn (061/22; 09.04.2022). A data protection concept ensures adherence to data protection regulations for the handled data. The dissemination strategies include discussing the results with the cancer counselling centres.Trial registration number German Clinical Trials Register (DRKS00028121); Pre-results

Specific N-Glycans Direct Apical Delivery of Transmembrane, but Not Soluble or Glycosylphosphatidylinositol-anchored Forms of Endolyn in Madin-Darby Canine Kidney Cells

The sialomucin endolyn is a transmembrane protein with a unique trafficking pattern in polarized Madin-Darby canine kidney cells. Despite the presence of a cytoplasmic tyrosine motif that, in isolation, is sufficient to mediate basolateral sorting of a reporter protein, endolyn predominantly traverses the apical surface en route to lysosomes. Apical delivery of endolyn is disrupted in tunicamycin-treated cells, implicating a role for N-glycosylation in apical sorting. Site-directed mutagenesis of endolyn's eight N-glycosylation sites was used to identify two N-glycans that seem to be the major determinants for efficient apical sorting of the protein. In addition, apical delivery of endolyn was disrupted when terminal processing of N-glycans was blocked using glycosidase inhibitors. Missorting of endolyn occurred independently of the presence or absence of the basolateral sorting signal, because apical delivery was also inhibited by tunicamycin when the cytoplasmic tyrosine motif was mutated. However, we found that apical secretion of a soluble mutant of endolyn was N-glycan independent, as was delivery of glycosylphosphatidylinositol-anchored endolyn. Thus, specific N-glycans are only essential for the apical sorting of transmembrane endolyn, suggesting fundamental differences in the mechanisms by which soluble, glycosylphosphatidylinositol-anchored, and transmembrane proteins are sorted

Vascular Targeting in Pancreatic Cancer: The Novel Tubulin-Binding Agent ZD6126 Reveals Antitumor Activity in Primary and Metastatic Tumor Models

ZD6126 is a novel vascular-targeting agent that acts by disrupting the tubulin cytoskeleton of an immature tumor endothelium, leading to an occlusion of tumor blood vessels and a subsequent tumor necrosis. We wanted to evaluate ZD6126 in primary and metastatic tumor models of human pancreatic cancer. Nude mice were injected orthotopically with L3.6pl pancreatic cancer cells. In single and multiple dosing experiments, mice received ZD6126, gemcitabine, a combination of both agents, or no treatment. For the induction of metastatic disease, additional groups of mice were injected with L3.6pl cells into the spleen. Twenty-four hours after a single-dose treatment, ZD6126 therapy led to an extensive central tumor necrosis, which was not seen after gemcitabine treatment. Multiple dosing of ZD6126 resulted in a significant growth inhibition of primary tumors and a marked reduction of spontaneous liver and lymph node metastases. Experimental metastatic disease could be significantly controlled by a combination of ZD6126 and gemcitabine, as shown by a reduction of the number and size of established liver metastases. As shown by additional in vitro and in vivo experiments, possible mechanisms involve antivascular activities and subsequent antiproliferative and proapoptotic effects of ZD6126 on tumor cells, whereas direct activities against tumor cells seem unlikely. These data highlight the antitumor and antimetastatic effects of ZD6126 in human pancreatic cancer and reveal benefits of adding ZD6126 to standard gemcitabine therapy

Men’s access to outpatient psychosocial cancer counseling

Hintergrund: Männer nehmen seltener als Frauen ambulante Krebsberatung in Anspruch, auch wenn sie davon profitieren könnten. Methode: In einer cluster-randomisierten Studie (registriert unter DRKS00032181) wurde geprüft, ob durch Maßnahmen auf verschiedenen Ebenen (Informierung von Zuweisenden, Öffentlichkeitsarbeit, strukturelle Veränderungen, männerspezifische Angebote) in einem Zeitraum von 12 Monaten der Anteil männlicher Ratsuchender in Krebsberatungsstellen erhöht werden kann (primärer Endpunkt: Erstkontakte, sekundärer Endpunkt: alle Kontakte). Der Interventionseffekt wurde durch Anpassen verallgemeinerter linearer gemischter Modelle als Odds Ratio ermittelt, welches für die Cluster-Struktur und den Anteil von Erst- beziehungsweise allen Kontakten während der 12 Monate vor Beginn der Intervention adjustiert ist. Ergebnisse: In 12 Regionen Deutschlands (je 6 im Interventions- beziehungsweise Kontrollarm) wurden 11 986 Personen erstmals beraten (Erstkontakte), davon 6 004 während der Interventionsphase. Der Männeranteil lag in diesem Zeitraum bei 30,7 % im Interventions- und 25,7 % im Kontrollarm. Das entspricht einem nicht signifikanten modellbasierten adjustierten Odds Ratio (OR) von 1,2 (95-%-Konfidenzintervall: [1,0; 1,4] p = 0,08) für den primären Endpunkt. Insgesamt 51 842 Beratungen (Erst- und Folgekontakte zusammen) wurden durchgeführt, 26 651 davon in der Interventionsphase. Der Männeranteil lag bei 27,6 % im Interventionsarm und 22,2 % im Kontrollarm. Das adjustierte OR für den sekundären Endpunkt ist 1,3 [1,1; 1,6] p = 0,01). Schlussfolgerung: Durch gezielte Umsetzung männerspezifischer Maßnahmen auf verschiedenen Ebenen kann der Männeranteil von Ratsuchenden in Krebsberatungsstellen leicht erhöht werden

[Specific Contribution of Psychosocial Cancer Counselling Centers - Perspectives of the Person Seeking Advice and the Referring Professional

Xyländer M, Bayer O, Schranz M, et al. Zum spezifischen Versorgungsbeitrag psychosozialer Krebsberatungsstellen – Sichtweisen der Ratsuchenden und Zuweisenden. Psychotherapie, Psychosomatik, medizinische Psychologie. 2021.AIM: The aim of the study is to look at the specific contribution of outpatient cancer counselling centers (OCCC) from the perspective of both the person seeking advice and the referring health care professionals.; METHODS: Qualitative design by means of guideline-based face-to-face interviews with cancer patients/relatives and individual telephone interviews with referring health care professionals.; RESULTS: A total of 43 persons seeking advice and 30 referring health care professionals were interviewed. With regard to the contents of counselling, psycho-oncological support and help for self-help in combination with social-legal information about additional support services are perceived as central features. In the group of referring physicians, however, there seems to be some uncertainty about what OCCCs (can) provide.; CONCLUSION: On the one hand, the results point to a specific core of the services offered by OCCCs, and on the other hand to ambiguous perceptions on the part of the respondents. They may contribute to further sharpening the profile of OCCC and to clarifying their place in the health care system. Thieme. All rights reserved

Pathways to cancer counselling centres: How do people become aware of them? A qualitative study with people seeking advice and those referring them

Schranz M, Bayer O, Xylaender M, et al. Wege in Krebsberatungsstellen: Wie werden Menschen darauf aufmerksam? Eine qualitative Studie mit Ratsuchenden und Zuweisenden. Onkologe. 2022.Background Outpatient cancer counselling centres (OCCC; in German Krebsberatungsstellen [KBS]) offer psychological, social and sociolegal support for people with cancer and their relatives, who are confronted not only with physical but also with psychosocial stress as a result of the disease. Research question How do people become aware of outpatient cancer counselling centres? Methods In a nationwide study, guided qualitative interviews were conducted with people seeking advice (cancer patients and relatives) and referrers (health care providers who refer to OCCC). The interviews with those seeking advice were conducted face-to-face and those with referrers by telephone. The data were analysed using content analysis according to Mayring. Results In all, 43 people seeking advice and 30 referrers were interviewed. A total of 31 h of audio material was recorded. From the respondents' point of view, flyers, information brochures and internet platforms are important sources for making patients and their relatives aware of OCCC. In addition to public relations, the personal approach in the clinical and non-clinical care system and in the social setting is of central importance. Some referrers address OCCC routinely, while others only do so when patients or relatives explicitly express a need for counselling. Conclusions In the medical sector, more attention could and should be drawn to OCCC as a helpful outpatient counselling setting. Closer cooperation between OCCC and general practitioners, oncologists and psycho-oncology services should be sought. Targeted and needs-oriented information can draw attention to the support offered by OCCC and thus improve the care of cancer patients.Hintergrund Ambulante Krebsberatungsstellen (KBS) bieten psychologische, soziale und sozialrechtliche Unterstützung für an Krebs erkrankte Menschen und Angehörige an, die infolge der Erkrankung nicht nur mit körperlichen, sondern auch mit psychosozialen Belastungen konfrontiert werden. Fragestellung Wie werden Menschen auf ambulante Krebsberatungsstellen aufmerksam? Methoden In einer deutschlandweiten Studie wurden leitfadengestützte qualitative Interviews mit Ratsuchenden (Krebspatient/innen und Angehörige) und Zuweisenden (Gesundheitsversorger/innen, die an KBS verweisen) geführt. Die Befragungen der Ratsuchenden erfolgten im persönlichen Gespräch, die der Zuweisenden telefonisch. Ausgewertet wurde inhaltsanalytisch nach Mayring. Ergebnisse Es wurden 43 Ratsuchende und 30 Zuweisende befragt. Insgesamt entstanden 31 h Audiomaterial. Aus Sicht der Befragten sind Flyer, Infobroschüren und Internetplattformen wichtige Quellen, um Patient/innen und deren Angehörige auf KBS aufmerksam zu machen. Neben der Öffentlichkeitsarbeit ist die persönliche Ansprache im klinischen wie nicht-klinischen Versorgungssystem und im sozialen Umfeld von zentraler Bedeutung. Manche Zuweisenden sprechen KBS routinemäßig an, andere erst bei explizit von Patient/innen oder Angehörigen geäußertem Beratungsbedarf. Schlussfolgerungen Im medizinischen Sektor könnte und sollte noch mehr auf KBS als hilfreiches ambulantes Beratungssetting hingewiesen werden. Eine engere Zusammenarbeit der KBS mit Hausärzt/innen, Onkolog/innen und psychoonkologischen Diensten ist anzustreben. Gezielte und bedarfsgerechte Informationen können auf das Unterstützungsangebot durch eine KBS hinweisen und so die Versorgung von Krebspatient/innen verbessern