Improving dose calculations on tomotherapy MVCT images.

The purpose of this investigation was the creation of a new protocol allowing more precise dose calculations on megavoltage CT (MVCT) images for tomotherapy, both for adaptive and StatRT planning. Daily MVCT images offer, next to positioning purposes, the possibility for daily dose check and adaptive planning. Dose calculations use the image value to density table (IVDT) to calculate physical densities from Hounsfield Units (HUs). These measured HUs change over time, leading to a dose calculation error. We noticed dose calculation variations due to IVDT changes of: 0.2% dose during a day, up to 1.6% dose from long-term variations, and up to 1.5% dose due to technical interventions. An analysis was performed applying the general methodology of a calibration problem. A model HU = bρc - 1020 was obtained using a weighted least squares inverse prediction method (HU as function of density) taking into account the heteroscedasticity. The b parameter is the major variable and depends also on the dose rate (DR). We demonstrate the correction for DR variations and the constance of the c parameter. Instead of scanning the whole tissue characterization phantom daily, we propose a simplified daily protocol: (a) morning airscan-like procedure with only two inserts on the table (defining b and thus the IVDT curve), (b) DR variations throughout the day can be corrected for using the DR model. A patient-specific protocol for which two inserts next to the patient are scanned could also be used, but results in equal uncertainties and is less practical. Therefore we recommend the morning procedure with dose rate variation correction. Applying the proposed transformations and the model, the correct IVDT of the moment can be reconstructed, with a simple measurement in the morning, and corrected with DR changes during the day. This corresponds with a linear mapping in time of the proposed IVDT function. The dosimetric variation is hereby reduced from up to 3% to 0.4 % for the tested pelvic and head-and-neck cases. In practice, several IVDT curves corresponding to "b" values can be entered. The correct IVDT curve of that moment can then be chosen from the list. Instead of the two high-density inserts on table, any calibrated single density phantom could be used in order to create the IVDT curve of the day, but it should have a larger size than the current inserts.

Alternative Splicing in Cancer and Immune Cells.

Splicing is a phenomenon enabling the excision of introns from pre-mRNA to give rise to mature mRNA. All the 20,000 genes of the human genome are concerned by this mechanism. Nevertheless, it is estimated that the proteome is composed of more than 100,000 proteins. How to go from 20,000 genes to more than 100,000 proteins? Alternative splicing (AS) is in charge of this diversity of proteins. AS which is found in most of the cells of an organism, participates in normal cells and in particular in immune cells, in the regulation of cellular behavior. In cancer, AS is highly dysregulated and involved in almost all of the hallmarks that characterize tumor cells. In view of the close link that exists between tumors and the immune system, we present in this review the literature relating to alternative splicing and immunotherapy. We also provide a global but not exhaustive view of AS in the immune system and tumor cells linked to the events that can lead to AS dysregulation in tumors.

Hematopoietic Prostaglandin D2 Synthase Controls Tfh/Th2 Communication and Limits Tfh Antitumor Effects.

T follicular helper (Tfh) cells are a subset of CD4+ T cells essential in immunity and have a role in helping B cells produce antibodies against pathogens. However, their role during cancer progression remains unknown. The mechanism of action of Tfh cells remains elusive because contradictory data have been reported on their protumor or antitumor responses in human and murine tumors. Like Tfh cells, Th2 cells are also involved in humoral immunity and are regularly associated with tumor progression and poor prognosis, mainly through their secretion of IL4. Here, we showed that Tfh cells expressed hematopoietic prostaglandin D2 (PGD2) synthase in a pSTAT1/pSTAT3-dependent manner. Tfh cells produced PGD2, which led to recruitment of Th2 cells via the PGD2 receptor chemoattractant receptor homologous molecule expressed on Th type 2 cells (CRTH2) and increased their effector functions. This cross-talk between Tfh and Th2 cells promoted IL4-dependent tumor growth. Correlation between Th2 cells, Tfh cells, and hematopoietic PGD2 synthase was observed in different human cancers and associated with outcome. This study provides evidence that Tfh/Th2 cross-talk through PGD2 limits the antitumor effects of Tfh cells and, therefore, could serve as a therapeutic target.

The Tumor Microenvironment Impairs Th1 IFNγ Secretion through Alternative Splicing Modifications of Irf1 Pre-mRNA.

It is clearly established that the immune system can affect cancer response to therapy. However, the influence of the tumor microenvironment (TME) on immune cells is not completely understood. In this respect, alternative splicing is increasingly described to affect the immune system. Here, we showed that the TME, via a TGFß-dependent mechanism, increased alternative splicing events and induced the expression of an alternative isoform of the IRF1 transcription factor (IRF1Δ7) in Th1 cells. We found that the SFPQ splicing factor (splicing factor, proline- and glutamine-rich) was responsible for the IRF1Δ7 production. We also showed, in both mice and humans, that the IRF1 alternative isoform altered the full-length IRF1 transcriptional activity on the Il12rb1 promoter, resulting in decreased IFNγ secretion in Th1 cells. Thus, the IRF1Δ7 isoform was increased in the TME, and inhibiting IRF1Δ7 expression could potentiate Th1 antitumor responses.

Cleaved Caspase-3 Transcriptionally Regulates Angiogenesis-Promoting Chemotherapy Resistance.

Caspases are well known for their role in apoptosis. Recently, nonapoptotic roles of caspases have been identified, however, these noncanonical roles are not well documented and the mechanisms involved are not fully understood. Here, we studied the role of cleaved caspase-3 using human- and mouse-proficient caspase-3 cancer cell lines and human-deficient caspase-3 cancer cells. Cleaved caspase-3 functioned as a transcription factor and directly bound to DNA. A DNA-binding domain was identified in the small subunit of caspase-3 and an active conformation was essential for caspase-3 transcriptional activity. Caspase-3 DNA binding enhanced angiogenesis by upregulating the expression of proangiogenic genes and by activating pathways that promoted endothelial cell activation. Some proapoptotic genes were downregulated in caspase-3-proficient cells. Inhibiting caspase-3 increased the efficacy of chemotherapy and decreased spontaneous tumor development. These data highlight a novel nonapoptotic role of caspase-3 and suggest that cleaved caspase-3 could be a new therapeutic target in cancer. SIGNIFICANCE: These findings report a noncanonical function of caspase-3 by demonstrating its ability to transcriptionally regulate the VEGFR pathway.

A bioinspired, reusable, paper-based system for high-performance large-scale evaporation.

A bioinspired, reusable, paper-based gold-nanoparticle film is fabricated by depositing an as-prepared gold-nanoparticle thin film on airlaid paper. This paper-based system with enhanced surface roughness and low thermal conductivity exhibits increased efficiency of evaporation, scale-up potential, and proven reusability. It is also demonstrated to be potentially useful in seawater desalination.