The approval of immune checkpoint inhibitors in combination with specific diagnostic biomarkers presents new challenges to pathologists as tumor tissue needs to be tested for expression of programmed ...death-ligand 1 (PD-L1) for a variety of indications. As there is currently no requirement to use companion diagnostic assays for PD-L1 testing in Germany different clones are used in daily routine. While the correlation of staining results has been tested in various entities, there is no data for head and neck squamous cell carcinomas (HNSCC) so far.
We tested five different PD-L1 clones (SP263, SP142, E1L3N, 22-8, 22C3) on primary HNSCC tumor tissue of 75 patients in the form of tissue microarrays. Stainings of both immune and tumor cells were then assessed and quantified by pathologists to simulate real-world routine diagnostics. The results were analyzed descriptively and the resulting staining pattern across patients was further investigated by principal component analysis and non-negative matrix factorization clustering.
Percentages of positive immune and tumor cells varied greatly. Both the resulting combined positive score as well as the eligibility for certain checkpoint inhibitor regimens was therefore strongly dependent on the choice of the antibody. No relevant co-clustering and low similarity of relative staining patterns across patients was found for the different antibodies.
Performance of different diagnostic anti PD-L1 antibody clones in HNSCC is less robust and interchangeable compared to reported data from other tumor entities. Determination of PD-L1 expression is critical for therapeutic decision making and may be aided by back-to-back testing of different PD-L1 clones.
Recently, a tumor cell-platelet interaction was identified in different tumor entities, resulting in a transfer of tumor-derived RNA into platelets, named further "tumor-educated platelets (TEP)". ...The present pilot study aims to investigate whether such a tumor-platelet transfer of RNA occurs also in patients suffering from head and neck squamous cell carcinoma (HNSCC).
Sequencing analysis of RNA derived from platelets of tumor patients (TPs) and healthy donors (HDs) were performed. Subsequently, quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used for verification of differentially expressed genes in platelets from TPs and HDs in a second cohort of patients and HDs. Data were analyzed by applying bioinformatic tools.
Sequencing of RNA derived from the tumor as well as from platelets of TPs and HDs revealed 426 significantly differentially existing RNA, at which 406 RNA were more and 20 RNA less abundant in platelets from TPs in comparison to that of HDs. In TPs' platelets, abundantly existing RNA coding for 49 genes were detected, characteristically expressed in epithelial cells and RNA, the products of which are involved in tumor progression. Applying bioinformatic tools and verification on a second TP/HD cohort, collagen type I alpha 1 chain (COL1A1) and zinc finger protein 750 (ZNF750) were identified as the strongest potentially platelet-RNA-sequencing (RNA-seq)-based biomarkers for HNSCC.
These results indicate a transfer of tumor-derived messenger RNA (mRNA) into platelets of HNSCC patients. Therefore, analyses of a patient's platelet RNA could be an efficient option for liquid biopsy in order to diagnose HNSCC or to monitor tumorigenesis as well as therapeutic responses at any time and in real time.
Btk and Vav proteins are all components of the signalosome that builds upon B cell receptor (BCR) activation. However, the role of Vav proteins within the signalosome is quite complex and not yet ...fully understood. Until now, studies of these have focused predominantly on a deficiency of Vav proteins alone or in combination with other Vav protein family members. Since a physical association of Btk with Vav was shown previously, we asked whether these molecules lie in the same or independent signaling pathways. By analyzing Vav1 and Vav3 single knock-out mice and generating double-knock-out animals deficient for either Vav1 or Vav3 and Btk, we observed, in line with previous publications, no severe B cell developmental defects when either Vav1 or Vav3 alone are not expressed. However, a simultaneous deficiency of Btk together with either Vav1 or Vav3 leads to a severe reduction of splenic B cells, which exhibit an immature phenotype. B cell developmental defects of Btk/Vav1-double deficient mice in the periphery were more severe than those observed in Btk-single-deficient animals. Additionally, morphological changes in splenic microarchitecture were observed in double- but also in single-knock-out mutants. These observations were accompanied by reduced BCR-induced Ca
mobilization, proliferation, germinal center formation and immunoglobulin secretion. Although deletion of Btk alone impaired Ca
mobilization upon BCR activation, the defect was even more severe when Vav1 or Vav3 were also mutated, indicating that Btk and the Vav proteins act in separate pathways that converge on Ca2+ signaling.
ASC differentiation suggests that both B and T cells contribute to the observed phenotype of a Btk/Vav-double deficiency. Our results show that Vav proteins and Btk are both components of the BCR-activated signalosome but control separate signaling pathways important for B cell development.
Abstract
Background
Head and neck squamous cell carcinomas (HNSCC) are highly immunosuppressive and aggressive malignancies. Exosomes are small extracellular vesicles that mediate intercellular ...communication. HNSCC patients have a significantly higher exosome load compared to healthy donors. Here, we evaluated the potential of plasma-derived exosomes as biomarkers for therapy monitoring in advanced HNSCC treated with conventional therapy.
Methods
Plasma from 17 HNSCC patients was collected before, during, and after treatment by surgery with adjuvant (chemo)radiation and at time of recurrence. Total exosomal protein (TEP) of exosomes was measured to estimate exosome load. Flow cytometry was used to evaluate relative fluorescence intensity (RFI) of tumor-associated and immunoregulatory proteins on exosomes. Effects of exosomes on CD8+ T cells and adenosine production by CD39+ T cells was assessed by flow cytometry and mass spectrometry.
Results
TEP levels and tumor-derived exosomes varied during and after therapy with an overall decrease in the tumor-free follow up but with an increase at time of recurrence. RFI values of immune checkpoints on exosomes, their potential for T cell inhibition, and adenosine production changed during and after therapy. Recurrent patients showed a different immunoinhibitory profile compared to disease-free patients. Exosomal PD-L1 emerged as the earliest discriminator for treatment failure and disease-free survival.
Conclusions
Plasma-derived exosomes from HNSCC patients emerge as promising non-invasive biomarkers for early detection of treatment failure. These exosomes can accurately inform about changes in patients’ immune status and may alert of impeding recurrence.
Zusammenfassung
Immuntherapeutische Agenzien haben mittlerweile einen festen Stellenwert in der Therapie verschiedenster Tumorentitäten inklusive der plattenepithelialen Kopf-Hals-Karzinome. Initial ...wurden sie im palliativen Setting eingesetzt, zunehmend aber auch in kurativer Intention, z. B. als Neoadjuvanz. In der heutigen Forschung geht es u. a. darum zu klären, welche Patienten von der Therapie profitieren und welche Kombinationstherapien erfolgreich sind. Im vorliegenden Artikel werden die relevanten Erkenntnisse der beiden internationalen Krebskongresse 2022 zusammengefasst.
High numbers of tumor-infiltrating lymphocytes (TIL) are linked to better survival in patients with cancer. Tissue-resident memory T cells (TRM; CD8+CD103+) are recognized as a key player of ...anticancer immune response. To assess TRM cells in primary, metastatic, and recurrent head and neck squamous cell carcinoma (HNSCC), we developed a tissue microarray (TMA) and used multiplex IHC (MxIHC).
Samples from primary tumors of 379 HNSCC cases treated at Southampton Hospitals between 2000 and 2016 were collected and analyzed. Of these, 105 cases had lymph node metastases and 82 recurrences. A TMA was generated with triplicate cores for each sample. MxIHC with a stain-and-strip approach was performed using CD8, CD103, and TIM3. Scanned slides were analyzed (digital image analysis) and quality checked (QC).
After QC, 194 primary tumors, 76 lymph node metastases, and 65 recurrences were evaluable. Alcohol consumption was statistically significantly correlated with a reduction of TRM cells in primary tumors (nondrinker vs. heavy drinker: P = 0.0036). The known survival benefit of TRM cell infiltration in primary tumors was not found for lymph node metastasis. In recurrences, a high TRM cell number led to a favorable outcome after 12 months. The checkpoint molecule TIM3, was expressed significantly higher on TRM and non-TRM cells in the lymph node compared with primary tumors (P < 0.0001), which was also seen in recurrences (P = 0.0134 and P = 0.0007, respectively).
We confirm the prognostic impact of TIL in primary tumors and in recurrences. TRM cell density in lymph node metastases was not linked to outcome. The role of TIM3, as a therapeutic target remains to be defined.
Background
Primary platinum‐based chemoradiotherapy (CRT) remains the treatment of choice for nonresectable squamous cell carcinoma of the head and neck (HNSCC). Immune‐checkpoint modulators are used ...as palliative therapy and studied in combination with definitive CRT. However, the immunological changes by CRT need yet to be understood.
Methods
A cohort consisting of 67 paired tissue biopsies (N = 134) of HNSCC patients before and after CRT was created. The expression of PD‐1, PD‐L1, and CD27 of tumor and immune cells by immunohistochemistry was evaluated.
Results
PD‐L1 expression on immune cells of non‐responders was significantly lower before CRT (P = .008). CD27 was expressed only on immune cells and not on cancer cells. A significant lower CD27‐expression score was observed following CRT (P = .019).
Conclusions
Conventional CRT changes the expression of CD27 in the tumor microenvironment. Whether this is due to a loss of expression or a reduction of CD27+ cells must be evaluated in further analyses.
Next to overall survival, quality of life is becoming more and more pivotal for cancer patients. The various domains of quality of life are complex and have different value to each patient. However, ...not only patients but also health care professionals, the pharmaceutical industry, and regulatory bodies ask: How can quality of life be reliably ascertained in clinical trials? For this purpose, carefully developed and validated specific questionnaires are needed: the patient-reported outcome measures (PROMs). A key challenge is to define how results based on PROMs can be used for shared decision-making. Next to clinical factors such as health and nutritional status, quality of life acts as a prognostic factor for overall survival in cancer. Thus, it is crucial to take quality of life into account in daily clinical practice.
Zusammenfassung
Neben dem möglichst langen Überleben spielt die Lebensqualität nach einer Krebserkrankung immer mehr eine Rolle für die Betroffenen. Die verschiedenen Lebensqualitätsbereiche sind ...komplex und werden individuell unterschiedlich gewichtet. Nicht nur für die Patient*innen, sondern auch für Ärzt*innen, die pharmazeutische Industrie und auch für die Zulassungsbehörden stellt sich die Frage: Wie kann man in klinischen Studien verlässlich feststellen, wie die Lebensqualität der Patient*innen ist? Dazu werden sorgfältig entwickelte und validierte Fragbögen benötigt, die Patient-Reported Outcome Measures (PROM). Die Herausforderung hierbei ist, Ergebnisse entsprechender Studien dazu nutzen zu können, die gemeinsame Entscheidungsfindung zu verbessern. Neben klinisch bekannten Faktoren wie Allgemein- und Ernährungszustand ist die Lebensqualität ebenfalls ein prognostischer Faktor für das Überleben im Rahmen von onkologischen Erkrankungen. Insofern spielt die Lebensqualität auch vor diesem Hintergrund eine klinisch bedeutsame Rolle und sollte entsprechend berücksichtigt werden.
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