The Nuclear Speckles Protein SRRM2 Is Exposed on the Surface of Cancer Cells.

The membrane composition of extracellular vesicles (EVs) largely reflects that of the plasma membrane of the cell of origin. We therefore hypothesized that EVs could be used for immunizations to generate monoclonal antibodies against well-known tumor antigens but possibly also against hitherto unknown tumor-associated target molecules. From an immunization experiment, we obtained a monoclonal antibody specific for SRRM2, an RNA-binding protein involved in splicing and a major component of nuclear speckles. Here, we used this antibody to demonstrate that SRRM2 is exposed on the surface of most cancer cell lines from various entities and, even more important, on cancer cells in vivo. Moreover, we demonstrated that SRRM2-specific CAR-T cells are functional in vitro and in vivo. Collectively, we identified SRRM2 as a promising new target molecule exposed on the cancer cell surface and showed that our SRRM2-specific antibody can be used as a basis for the development of new targeted cancer therapies.

Tumor Treating Fields (TTFields) Induce Cell Junction Alterations in a Human 3D In Vitro Model of the Blood-Brain Barrier.

In a recent study, we showed in an in vitro murine cerebellar microvascular endothelial cell (cerebEND) model as well as in vivo in rats that Tumor-Treating Fields (TTFields) reversibly open the blood-brain barrier (BBB). This process is facilitated by delocalizing tight junction proteins such as claudin-5 from the membrane to the cytoplasm. In investigating the possibility that the same effects could be observed in human-derived cells, a 3D co-culture model of the BBB was established consisting of primary microvascular brain endothelial cells (HBMVEC) and immortalized pericytes, both of human origin. The TTFields at a frequency of 100 kHz administered for 72 h increased the permeability of our human-derived BBB model. The integrity of the BBB had already recovered 48 h post-TTFields, which is earlier than that observed in cerebEND. The data presented herein validate the previously observed effects of TTFields in murine models. Moreover, due to the fact that human cell-based in vitro models more closely resemble patient-derived entities, our findings are highly relevant for pre-clinical studies.

Tumor Treating Fields (TTFields) Reversibly Permeabilize the Blood-Brain Barrier In Vitro and In Vivo.

Despite the availability of numerous therapeutic substances that could potentially target CNS disorders, an inability of these agents to cross the restrictive blood-brain barrier (BBB) limits their clinical utility. Novel strategies to overcome the BBB are therefore needed to improve drug delivery. We report, for the first time, how Tumor Treating Fields (TTFields), approved for glioblastoma (GBM), affect the BBB's integrity and permeability. Here, we treated murine microvascular cerebellar endothelial cells (cerebEND) with 100-300 kHz TTFields for up to 72 h and analyzed the expression of barrier proteins by immunofluorescence staining and Western blot. In vivo, compounds normally unable to cross the BBB were traced in healthy rat brain following TTFields administration at 100 kHz. The effects were analyzed via MRI and immunohistochemical staining of tight-junction proteins. Furthermore, GBM tumor-bearing rats were treated with paclitaxel (PTX), a chemotherapeutic normally restricted by the BBB combined with TTFields at 100 kHz. The tumor volume was reduced with TTFields plus PTX, relative to either treatment alone. In vitro, we demonstrate that TTFields transiently disrupted BBB function at 100 kHz through a Rho kinase-mediated tight junction claudin-5 phosphorylation pathway. Altogether, if translated into clinical use, TTFields could represent a novel CNS drug delivery strategy.

The importance of the pericardium for cardiac biomechanics: from physiology to computational modeling.

The human heart is enclosed in the pericardial cavity. The pericardium consists of a layered thin sac and is separated from the myocardium by a thin film of fluid. It provides a fixture in space and frictionless sliding of the myocardium. The influence of the pericardium is essential for predictive mechanical simulations of the heart. However, there is no consensus on physiologically correct and computationally tractable pericardial boundary conditions. Here, we propose to model the pericardial influence as a parallel spring and dashpot acting in normal direction to the epicardium. Using a four-chamber geometry, we compare a model with pericardial boundary conditions to a model with fixated apex. The influence of pericardial stiffness is demonstrated in a parametric study. Comparing simulation results to measurements from cine magnetic resonance imaging reveals that adding pericardial boundary conditions yields a better approximation with respect to atrioventricular plane displacement, atrial filling, and overall spatial approximation error. We demonstrate that this simple model of pericardial-myocardial interaction can correctly predict the pumping mechanisms of the heart as previously assessed in clinical studies. Utilizing a pericardial model not only can provide much more realistic cardiac mechanics simulations but also allows new insights into pericardial-myocardial interaction which cannot be assessed in clinical measurements yet.

Multiphysics Modeling of the Atrial Systole under Standard Ablation Strategies.

The aim of this study was to develop a computational framework to compare the impact of standard ablation concepts on the mechanical performance of the atria, since different line combinations cannot be applied in practice to the same patient. For this purpuse, we coupled electro-mechano-hemodynamic mathematical models based on biophysical principles and simulate the contractile performance of the atria. We computed systolic pressures and volumes in two patient-specific atrial geometries (one of normal size and one hypertrophied) with various ablation concepts. We found that our computational model is able to detect the differences in the left atrial contractility and ejection fraction for various electrical activation sequences resulting from different ablation line combinations. We show that multiphysics modeling has the potential to quantify the hemodynamic performance of left atria for different ablation lines, which could be used as additional pre-operative clinical information for the choice of the ablation concept in the future.