Objective MicroRNA (miRNA) expression profile can be used as prognostic marker for human cancers. We aim to explore the significance of miRNAs in colorectal cancer (CRC) metastasis. Design We performed miRNA microarrays using primary CRC tissues from patients with and without metastasis, and validated selected candidates in 85 CRC samples by quantitative real-time PCR (qRT-PCR). We tested metastatic activity of selected miRNAs and identified miRNA targets by prediction algorithms, qRT-PCR, western blot and luciferase assays. Clinical outcomes were analysed in six sets of CRC cases (n=449), including The Cancer Genome Atlas (TCGA) consortium and correlated with miR-224 status. We used the Kaplan–Meier method and log-rank test to assess the difference in survival between patients with low or high levels of miR-224 expression. Results MiR-224 expression increases consistently with tumour burden and microsatellite stable status, and miR-224 enhances CRC metastasis in vitro and in vivo. We identified SMAD4 as a miR-224 target and observed negative correlation (Spearman Rs=−0.44, p<0.0001) between SMAD4 and miR-224 expression in clinical samples. Patients with high miR-224 levels display shorter overall survival in multiple CRC cohorts (p=0.0259, 0.0137, 0.0207, 0.0181, 0.0331 and 0.0037, respectively), and shorter metastasis-free survival (HR 6.51, 95% CI 1.97 to 21.51, p=0.0008). In the TCGA set, combined analysis of miR-224 with SMAD4 expression enhanced correlation with survival (HR 4.12, 95% CI 1.1 to 15.41, p=0.0175). Conclusions MiR-224 promotes CRC metastasis, at least in part, through the regulation of SMAD4. MiR-224 expression in primary CRC, alone or combined with its targets, may have prognostic value for survival of patients with CRC.

FLASH radiation therapy (RT) is a promising new paradigm in radiation oncology. However, a major question that remains is the robustness and reproducibility of the FLASH effect when different irradiators are used on animals or patients with different genetic backgrounds, diets, and microbiomes, all of which can influence the effects of radiation on normal tissues. To address questions of rigor and reproducibility across different centers, we analyzed independent data sets from The University of Texas MD Anderson Cancer Center and from Lausanne University (CHUV). Both centers investigated acute effects after total abdominal irradiation to C57BL/6 animals delivered by the FLASH Mobetron system. The two centers used similar beam parameters but otherwise conducted the studies independently. The FLASH-enabled animal survival and intestinal crypt regeneration after irradiation were comparable between the two centers. These findings, together with previously published data using a converted linear accelerator, show that a robust and reproducible FLASH effect can be induced as long as the same set of irradiation parameters are used.

Sepsis remains a leading cause of death for humans and currently has no pathogenesis-specific therapy. Hampered progress is partly due to a lack of insight into deep mechanistic processes. In the past decade, deciphering the functions of small noncoding miRNAs in sepsis pathogenesis became a dynamic research topic. To screen for new miRNA targets for sepsis therapeutics, we used samples for miRNA array analysis of PBMCs from patients with sepsis and control individuals, blood samples from 2 cohorts of patients with sepsis, and multiple animal models: mouse cecum ligation puncture–induced (CLP-induced) sepsis, mouse viral miRNA challenge, and baboon Gram⁺ and Gram^– sepsis models. miR-93-5p met the criteria for a therapeutic target, as it was overexpressed in baboons that died early after induction of sepsis, was downregulated in patients who survived after sepsis, and correlated with negative clinical prognosticators for sepsis. Therapeutically, inhibition of miR-93-5p prolonged the overall survival of mice with CLP-induced sepsis, with a stronger effect in older mice. Mechanistically, anti–miR-93-5p therapy reduced inflammatory monocytes and increased circulating effector memory T cells, especially the CD4⁺ subset. AGO2 IP in miR-93–KO T cells identified important regulatory receptors, such as CD28, as direct miR-93-5p target genes. In conclusion, miR-93-5p is a potential therapeutic target in sepsis through the regulation of both innate and adaptive immunity, with possibly a greater benefit for elderly patients than for young patients.

Background The understanding of how varying radiation beam parameter settings affect the induction and magnitude of the FLASH effect remains limited. Purpose We sought to evaluate how the magnitude of radiation-induced gastrointestinal (GI) toxicity (RIGIT) depends on the interplay between mean dose rate (MDR) and dose per pulse (DPP). Methods C57BL/6J mice were subjected to total abdominal irradiation (11-14 Gy single fraction) under conventional irradiation (low DPP and low MDR, CONV) and various combinations of DPP and MDR up to ultra-high-dose-rate (UHDR) beam conditions. The effects of DPP were evaluated for DPPs of 1-6 Gy while the total dose and MDR were kept constant; the effects of MDR were evaluated for the range 0.3– 1440 Gy/s while the total dose and DPP were kept constant. RIGIT was quantified in non-tumor–bearing mice through the regenerating crypt assay and survival assessment. Tumor response was evaluated through tumor growth delay. Results Within each tested total dose using a constant MDR (>100 Gy/s), increasing DPP led to better sparing of regenerating crypts, with a more prominent effect seen at 12 and 14 Gy TAI. However, at fixed DPPs >4 Gy, similar sparing of crypts was demonstrated irrespective of MDR (from 0.3 to 1440 Gy/s). At a fixed high DPP of 4.7 Gy, survival was equivalently improved relative to CONV for all MDRs from 0.3 Gy/s to 104 Gy/s, but at a lower DPP of 0.93 Gy, increasing MDR produced a greater survival effect. We also confirmed that high DPP, regardless of MDR, produced the same magnitude of tumor growth delay relative to CONV using a clinically relevant melanoma mouse model. Conclusions This study demonstrates the strong influence that the beam parameter settings have on the magnitude of the FLASH effect. Both high DPP and UHDR appeared independently sufficient to produce FLASH sparing of GI toxicity, while isoeffective tumor response was maintained across all conditions.

Proper characterization of ¹³⁷Cs sources used for dosimeter calibration and performance testing is crucial for accurate and precise knowledge of air kerma rate and delivered dose. A 269 GBq ¹³⁷Cs source was relocated to a new facility, which had a footprint of approximately 2.8 m × 3.6 m and a 3.4 m high ceiling. A small room size, such as in the new facility, may significantly increase backscatter from the walls or room return. Due to the limited source strength, a relatively close irradiation position of 1.00 m from the source was selected to decrease required exposure times. Proximity to the small but cylindrical source has the potential to alter the 1/r ² relationship (inverse square law) of air kerma rate with distance associated with point sources. Practical tutorials concerning dosimetry irradiation facilities are largely absent from the archival literature. For those reasons, standard characterization experiments were repeated multiple times, great care was taken with positioning, several experiments were added to the standard ones, an ion chamber and dosimeters were used rather than film for greater accuracy and precision, and practical details of the process were recorded. The impacts of room size and finite source dimensions were quantified and mitigated by the efforts reported here. Recommendations for simple but thorough facility characterization, beam calibration, and quality control resulted.

A series of experiments were conducted using commercially available Al ₂ O ₃ :C optically stimulated luminescent dosimeters to provide a technical basis for their precise calibration and statistical performance at irradiated air kerma doses between 0.02 mGy and 5 mGy using ¹³⁷ Cs. This study examines the dose response linearity, studies the background signal for annealed dosimeters, and compares the statistical performance of dosimeters that were annealed and not annealed prior to their irradiation and readout. The average and standard deviation for the response of groups of dosimeters annealed and nonannealed prior to their irradiation were determined at each delivered dose. The batch of dosimeters that were annealed prior to their irradiation exhibited a coefficient of variation in its mean dose response below 10% when using three or more irradiation trials at each delivered air kerma dose between 0.02 mGy and 5 mGy. The reader calibration factor was calculated using the response of the annealed batch of dosimeters and was determined to be 756 ± 7 photomultiplier tube counts per mGy. Best estimates of the individual sensitivity factors were determined to be between 0.79 and 1.12 for the annealed batch of dosimeters. The minimum number of irradiations required to accurately determine the sensitivity factor of each individual dosimeter is reported with the recommended reader and dosimeter calibration procedures.