Head and neck cancer of unknown primary: unveiling primary tumor sites through machine learning on DNA methylation profiles.

BACKGROUND: The unknown tissue of origin in head and neck cancer of unknown primary (hnCUP) leads to invasive diagnostic procedures and unspecific and potentially inefficient treatment options for patients. The most common histologic subtype, squamous cell carcinoma, can stem from various tumor primary sites, including the oral cavity, oropharynx, larynx, head and neck skin, lungs, and esophagus. DNA methylation profiles are highly tissue-specific and have been successfully used to classify tissue origin. We therefore developed a support vector machine (SVM) classifier trained with publicly available DNA methylation profiles of commonly cervically metastasizing squamous cell carcinomas (n = 1103) in order to identify the primary tissue of origin of our own cohort of squamous cell hnCUP patient's samples (n = 28). Methylation analysis was performed with Infinium MethylationEPIC v1.0 BeadChip by Illumina. RESULTS: The SVM algorithm achieved the highest overall accuracy of tested classifiers, with 87%. Squamous cell hnCUP samples on DNA methylation level resembled squamous cell carcinomas commonly metastasizing into cervical lymph nodes. The most frequently predicted cancer localization was the oral cavity in 11 cases (39%), followed by the oropharynx and larynx (both 7, 25%), skin (2, 7%), and esophagus (1, 4%). These frequencies concord with the expected distribution of lymph node metastases in epidemiological studies. CONCLUSIONS: On DNA methylation level, hnCUP is comparable to primary tumor tissue cancer types that commonly metastasize to cervical lymph nodes. Our SVM-based classifier can accurately predict these cancers' tissues of origin and could significantly reduce the invasiveness of hnCUP diagnostics and enable a more precise therapy after clinical validation.

Hsp70-A Universal Biomarker for Predicting Therapeutic Failure in Human Female Cancers and a Target for CTC Isolation in Advanced Cancers.

Heat shock protein 70 (Hsp70) is frequently overexpressed in many different tumor types. However, Hsp70 has also been shown to be selectively presented on the plasma membrane of tumor cells, but not normal cells, and this membrane form of Hsp70 (mHsp70) could be considered a universal tumor biomarker. Since viable, mHsp70-positive tumor cells actively release Hsp70 in lipid micro-vesicles, we investigated the utility of Hsp70 in circulation as a universal tumor biomarker and its potential as an early predictive marker of therapeutic failure. We have also evaluated mHsp70 as a target for the isolation and enumeration of circulating tumor cells (CTCs) in patients with different tumor entities. Circulating vesicular Hsp70 levels were measured in the peripheral blood of tumor patients with the compHsp70 ELISA. CTCs were isolated using cmHsp70.1 and EpCAM monoclonal antibody (mAb)-based bead approaches and characterized by immunohistochemistry using cytokeratin and CD45-specific antibodies. In two out of 35 patients exhibiting therapeutic failure two years after initial diagnosis of non-metastatic breast cancer, progressively increasing levels of circulating Hsp70 had already been observed during therapy, whereas levels in patients without subsequent recurrence remained unaltered. With regards to CTC isolation from patients with different tumors, an Hsp70 mAb-based selection system appears superior to an EpCAM mAb-based approach. Extracellular and mHsp70 can therefore serve as a predictive biomarker for therapeutic failure in early-stage tumors and as a target for the isolation of CTCs in various tumor diseases.

How to... train your skills goes digital! A project report on the development and implementation of practice-oriented digital student tutorials.

Objective: This project report outlines the development and implementation of digital tutorials ("How to... train your skills goes digital!") by peer tutors (TUTs) based on previously established in-person tutorials, as well as the subsequent combination of both approaches. The TUTs' objective, in spite of corona-related restrictions and strict hygiene requirements during the pandemic, was to provide fellow students with the opportunity to learn, practice and reflect on clinical-practical skills. Methodology: In a collaborative undertaking, the TUTs first analyzed the learning objectives of the in-person tutorials in order to be able to design content-matched digital tutorials without entirely abandoning the practical aspect. The Moodle learning management platform was selected as the appropriate tool for delivery of the relevant theoretical knowledge. Practical exercises are embedded in the digital tutorials during online meetings. The participants (PTs) create their own models in the sense of a home skills station based on instructions provided via Moodle. Acceptance was systematically documented via questionnaires. Results: The digital tutorials were well accepted by the PTs (n=64). Evaluation (response rate: 37.5%) outcomes were consistently positive. Both course implementation and the PTs' own progress were rated "good" to "very good". Nevertheless, the PTs do not yet feel well-prepared to carry out the various activities practiced independently. In the winter semester of 2020/21, the in-person tutorials were also reintroduced in a combined format. The marked demand for the tutorials may indicate the PT preference for practice on the simulation center models. Conclusion: The systematic combination of digital and in-person tutorials using the flipped classroom approach would appear to make sense in the long run. The effectiveness and sustainability of this approach in comparison with in-person tutorials only should be further investigated.