Circulating Hsp70 Levels and the Immunophenotype of Peripheral Blood Lymphocytes as Potential Biomarkers for Advanced Lung Cancer and Therapy Failure after Surgery.

Lung cancer remains a devastating disease with a poor clinical outcome. A biomarker signature which could distinguish lung cancer from metastatic disease and detect therapeutic failure would significantly improve patient management and allow for individualized, risk-adjusted therapeutic decisions. In this study, circulating Hsp70 levels were measured using ELISA, and the immunophenotype of the peripheral blood lymphocytes were measured using multiparameter flow cytometry, to identify a predictive biomarker signature for lung cancer patients pre- and post-operatively, in patients with lung metastases and in patients with COPD as an inflammatory lung disease. The lowest Hsp70 concentrations were found in the healthy controls followed by the patients with advanced COPD. Hsp70 levels sequentially increased with an advancing tumor stage and metastatic disease. In the early-recurrence patients, Hsp70 levels started to increase within the first three months after surgery, but remained unaltered in the recurrence-free patients. An early recurrence was associated with a significant drop in B cells and an increase in Tregs, whereas the recurrence-free patients had elevated T and NK cell levels. We conclude that circulating Hsp70 concentrations might have the potential to distinguish lung cancer from metastatic disease, and might be able to predict an advanced tumor stage and early recurrence in lung cancer patients. Further studies with larger patient cohorts and longer follow-up periods are needed to validate Hsp70 and immunophenotypic profiles as predictive biomarker signatures.

Anastomotic Rings and Inflammation Values as Biomarkers for Leakage of Stapled Circular Colorectal Anastomoses.

Reliable markers to predict or diagnose anastomotic leakage (AL) of stapled circular anastomoses following colorectal resections are an important clinical need. Here, we aim to quantitatively investigate the morphology of anastomotic rings as an early available prognostic marker for AL and compare them to established inflammatory markers. We perform a prospective single-center cohort study, including patients undergoing stapled circular anastomosis between August 2020 and August 2021. The predictive value of the anastomotic ring configuration and the neutrophil-to-lymphocyte ratio (NLR) regarding anastomotic leakage is examined by ROC analyses and compared to the C-reactive protein (CRP) as an established marker. We included 204 patients, of which 19 suffered from anastomotic leakage (LEAK group), while in 185 patients the anastomoses healed well (HEAL group). The minimal height of the anastomotic rings as a binary classifier had a good ROC-AUC of 0.81 but was inferior to the NLR at postoperative day (POD) 5, with an excellent ROC-AUC of 0.93. Still, it was superior to the NLR at POD 3 (0.74) and the CRP at POD 3 (ROC-AUC 0.54) and 5 (ROC-AUC 0.70). The minimal height of the anastomotic rings as indicator for technically insufficient anastomoses is a good predictor of AL, while postoperatively the NLR was superior to the CRP in prediction of AL.

Bioluminescence Imaging in the Chick Chorioallantoic Membrane Assay.

For the analysis of tumorigenesis and therapeutic intervention, high throughput technologies that allow the detection of tumor size in the context of a living organism are of need. Here we describe the use of a chorioallantoic membrane model in the developing chick embryo on which growth of a tumor xenograft can be monitored over time, enabling bioluminescence technology.

Non-invasive imaging of engineered human tumors in the living chicken embryo.

The growing interest in engineered tumor models prompted us to devise a method for the non-invasive assessment of such models. Here, we report on bioluminescence imaging (BLI) for the assessment of engineered tumor models in the fertilized chicken egg, i.e, chick chorioallantoic membrane (CAM) assay. One prostate cancer (PC-3) and two osteosarcoma (MG63 and HOS) cell lines were modified with luciferase reporter genes. To create engineered tumors, these cell lines were seeded either onto basement membrane extract (BME) or gelfoam scaffolds, and subsequently grafted in vivo onto the CAM. BLI enabled non-invasive, specific detection of the engineered tumors on the CAM in the living chicken embryo. Further, BLI permitted daily, quantitative monitoring of the engineered tumors over the course of up to 7 days. Data showed that an extracellular matrix (ECM) composed of BME supported growth of reporter gene marked PC-3 tumors but did not support MG63 or HOS tumor growth. However, MG63 tumors engineered on the collagen-based gelfoam ECM showed a temporal proliferation burst in MG63 tumors. Together, the data demonstrated imaging of engineered human cancer models in living chicken embryos. The combination of CAM assay and BLI holds significant potential for the examination of a broad range of engineered tumor models.