Ultrashort pulse infrared lasers can simultaneously sample and homogenize biological tissue using desorption by impulsive vibrational excitation (DIVE). With growing attention on alterations in lipid metabolism in malignant disease, mass spectrometry (MS)-based lipidomic analysis has become an emerging topic in cancer research. In this pilot study, we investigated the feasibility of tissue sampling with a nanosecond infrared laser (NIRL) for the subsequent lipidomic analysis of oropharyngeal tissues, and its potential to discriminate oropharyngeal squamous cell carcinoma (OPSCC) from non-tumorous oropharyngeal tissue. Eleven fresh frozen oropharyngeal tissue samples were ablated. The produced aerosols were collected by a glass fiber filter, and the lipidomes were analyzed with mass spectrometry. Data was evaluated by principal component analysis and Welch’s t-tests. Lipid profiles comprised 13 lipid classes and up to 755 lipid species. We found significant inter- and intrapatient alterations in lipid profiles for tumor and non-tumor samples (p-value < 0.05, two-fold difference). Thus, NIRL tissue sampling with consecutive MS lipidomic analysis is a feasible and promising approach for the differentiation of OPSCC and non-tumorous oropharyngeal tissue and may provide new insights into lipid composition alterations in OPSCC.

Die ORATOR2-Studie war eine internationale, multizentrische, randomisierte Phase-II-Studie mit dem Ziel, das optimale Therapie-Deeskalationskonzept bei HPV-positiven Oropharynxkarzinomen im Frühstadium (T1–2, N0–2) zu erfassen. Die Patienten (n = 61) wurden entweder mittels transoraler Chirurgie und Neck Dissection behandelt oder mittels intensitätsmodulierter Strahlentherapie. Die Deeskalationskomponente beider Arme war eine Dosisreduktion der primären oder adjuvanten Strahlentherapie. Der primäre Endpunkt war das Gesamtüberleben. Dieses war im Bestrahlungsarm besser, was hauptsächlich durch 3 Todesfälle im Operationsarm bedingt war, hiervon 2 therapieassoziiert. Dies führte zum frühzeitigen Abbruch der Studie. Die Deutsche Gesellschaft für Hals-Nasen-Ohren-Heilkunde, Kopf- und Hals-Chirurgie/Arbeitsgemeinschaft für Onkologie warnt davor, die Studienergebnisse als mögliches Entscheidungskriterium für die Therapiewahl von HPV-positiven Oropharynxkarzinomen zu verwenden, angesichts der Schwächen der Studie insbesondere im Operationsarm (Sicherheitsabstand von 10 mm, streng empfohlene Nachresektion bei R1/knappen Resektionsrändern, keine freien/regionalen Transplantate zur Rekonstruktion, kaum Verwendung der Laserchirurgie, hohe Tracheotomierate). Kleine Patientenzahlen, eine sehr selektionierte Patientenkohorte und die kurze Nachbeobachtungszeit schränken die Aussagekraft der Studie weiter ein. Patienten mit HPV-assoziierten Oropharynxkarzinomen sollten aktuell keine deeskalierende (Strahlen-) Therapie außerhalb klinischer Studien erhalten. Bei der Entscheidung zwischen einem chirurgischen oder einem strahlentherapeutischen Vorgehen sollten Patienten nach interdisziplinärer Zustimmung eines Tumorboards über Vor- und Nachteile beider Modalitäten aufgeklärt werden, bis Ergebnisse klinisch relevanter Phase-III-Studien (bspw. EORTC 1420) vorliegen. ORATOR2 was a randomized phase II trial aiming to assess an optimal approach for therapy de-escalation in early (T1-T2, N0-N2) human papillomavirus (HPV)-related oropharyngeal squamous cell carcinomas (OPSCC). Radiotherapy (RT) (consisting of a reduced dose of 60 Gy with concurrent weekly cisplatin in N+ patients) was compared to trans-oral surgery (TOS) and neck dissection (ND) (with adjuvant reduced-dose RT depending on pathologic findings) in 61 patients. The primary endpoint, overall survival, favored the radiotherapy approach. This was mainly due to 3 mortality events in the surgery arm (2 surgery-related) which resulted in an early trial termination. The authors, who speak on behalf of the German Society of Otorhinolaryngology, Head & Neck Surgery (working group for oncology) warn to draw conclusions for clinical practice pointing out the main shortages/weaknesses of this trial especially in the surgery arm (at least 1 cm margins, recommending re-operation if not achieved, prohibition of regional or free flaps, high rates of tracheotomy, low rate of TLM). Small patient numbers, a highly selected patient cohort and a short follow-up time further limit this study’s relevance. Therefore, patients with HPV-related OPSCC should not receive de-escalating (radiation) therapy outside of clinical trials. When deciding between a surgical or a radio-therapeutical approach, patients should be informed about the pros and cons of both modalities after interdisciplinary consent in a tumor board, as long as clinical trial results` (e. g. EORTC 1420) are pending. Article published online: 01 March 2023 © 2023. Thieme. All rights reserved. Georg Thieme Verlag KG Rüdigerstraße 14, 70469 Stuttgart, Germany

Background/Objectives: Human papillomavirus (HPV) infection is a well-established risk factor for oropharyngeal squamous cell carcinoma (OPSCC), where p16 immunohistochemistry serves as a surrogate marker. However, the role of HPV in sinonasal squamous cell carcinoma (SNSCC) remains less defined, and the reliability of p16 as a standalone surrogate is under debate. This study aimed to assess the concordance between p16 expression and HPV-DNA status in SNSCC and characterize clinicopathologic features in HPV-associated cases. Methods: We retrospectively analyzed 111 SNSCC cases diagnosed between 2008 and 2024 at two German centers. p16 status was determined by immunohistochemistry using site-specific antibody protocols. HPV-DNA testing and genotyping were performed via PCR and reverse hybridization. Clinical and histopathological data were collected and compared between HPV-positive and -negative tumors. Results: HPV-DNA was detected in 31/111 cases (27.9%), with HPV16 and HPV33 (Site A) and HPV 16 and HPV18 (Site B) being the most frequent subtypes. Discordance between p16 and HPV-DNA status was observed in 29.7% of cases, with site-specific discordance rates of 44.6% and 14.5%. Patients with HPV-positive tumors were younger than their HPV-negative counterparts. Conclusions: Our findings underscore the limitations of p16 as a single surrogate marker for detecting HPV-associated sinonasal cancer. Future research on the role of HPV in sinonasal cancer should integrate complementary testing methods (like p16Ink4A immunohistochemistry and HPV DNA/mRNA analysis) and aim for test standardization.