Targeting AKT/mTOR pathway in order to sensitise rectal tumour cells to irradiation using patient derived organoids as a model

Treatment resistance is one of the key contributors to low overall survival rates in Colorectal cancer (CRC) patients. Although both resistance to chemotherapy and radiotherapy is observed in CRC patients, radioresistance of rectal cancer patients is a major issue in the clinic. 40% of patients with rectal cancer that receive neo-adjuvant radiotherapy will have no significant response to treatment, whereas only 10% of patients will have a complete pathological response (pathCR) to treatment. In order to understand pathCR radiosensitivity markers need to be defined. Studying radiotherapy with current models has been challenging due to the nature of the available models and their limitations to recapitulate the patients’ tumour biology. The emerging new 3 dimensional (3D) organoid models show promise in regards to the modelling of cancer and patients’ response with in vitro experiments that will be able to recapitulate tumour microenvironment. The aim of this study was to establish an organoid model for studying irradiation in rectal cancer patients and to identify radiosensitivity and radioresistance drivers in order to understand and tackle the problem of resistance to radiotherapy in clinical patients. Organoid lines were established and characterised for pathological and molecular features. The organoids were subjected to short-course radiotherapy (25Gy dose delivered in 5 fractions over the course of 5 days) and the response to the treatment was measured. Furthermore, the changes caused by the irradiation were investigated by performing whole genome sequencing, DNA methylation arrays, total RNA sequencing, and single-cell sequencing on irradiated and control organoids. Lastly, combination drug and radiotherapy assays were performed using organoids with mTOR and Akt inhibitors in order to sensitise cells to irradiation. The results revealed that irradiation causes changes on a genome-wide scale and disrupts the mTOR/PiK3CA signalling pathway. Combination therapy showed that Rapamycin is not effective in sensitising cells to irradiation, whereas AZD2014 was able to sensitise certain organoid lines to irradiation; the same was found to be the case for MK-2206. In conclusion, the results showed organoids pose as representative models for modelling radiotherapy response, and that blocking mTOR via dual inhibition of mTORC1 and mTORC2 as well as inhibition of Akt can sensitise cells to irradiation. Finally it was found that, in order to sensitise resistant lines, the dual inhibition of mTOR/Akt might be required

Neutron-rich Chromium Isotope Anomalies in Supernova Nanoparticles

Neutron-rich isotopes with masses near that of iron are produced in Type Ia and II supernovae (SNeIa and SNeII). Traces of such nucleosynthesis are found in primitive meteorites in the form of variations in the isotopic abundance of ^(54)Cr, the most neutron-rich stable isotope of chromium. The hosts of these isotopic anomalies must be presolar grains that condensed in the outflows of SNe, offering the opportunity to study the nucleosynthesis of iron-peak nuclei in ways that complement spectroscopic observations and can inform models of stellar evolution. However, despite almost two decades of extensive search, the carrier of ^(54)Cr anomalies is still unknown, presumably because it is fine grained and is chemically labile. Here, we identify in the primitive meteorite Orgueil the carrier of ^(54)Cr anomalies as nanoparticles (3.6 × solar). Such large enrichments in ^(54)Cr can only be produced in SNe. The mineralogy of the grains supports condensation in the O/Ne-O/C zones of an SNII, although a Type Ia origin cannot be excluded. We suggest that planetary materials incorporated different amounts of these nanoparticles, possibly due to late injection by a nearby SN that also delivered ^(26)Al and ^(60)Fe to the solar system. This idea explains why the relative abundance of ^(54)Cr and other neutron-rich isotopes vary between planets and meteorites. We anticipate that future isotopic studies of the grains identified here will shed new light on the birth of the solar system and the conditions in SNe

Ion distribution and ablation depth measurements of a fs-ps laser-irradiated solid tin target

The ablation of solid tin surfaces by an 800-nanometer-wavelength laser is studied for a pulse length range from 500 fs to 4.5 ps and a fluence range spanning 0.9 to 22 J/cm^2. The ablation depth and volume are obtained employing a high-numerical-aperture optical microscope, while the ion yield and energy distributions are obtained from a set of Faraday cups set up under various angles. We found a slight increase of the ion yield for an increasing pulse length, while the ablation depth is slightly decreasing. The ablation volume remained constant as a function of pulse length. The ablation depth follows a two-region logarithmic dependence on the fluence, in agreement with the available literature and theory. In the examined fluence range, the ion yield angular distribution is sharply peaked along the target normal at low fluences but rapidly broadens with increasing fluence. The total ionization fraction increases monotonically with fluence to a 5-6% maximum, which is substantially lower than the typical ionization fractions obtained with nanosecond-pulse ablation. The angular distribution of the ions does not depend on the laser pulse length within the measurement uncertainty. These results are of particular interest for the possible utilization of fs-ps laser systems in plasma sources of extreme ultraviolet light for nanolithography.Comment: 8 pages, 7 figure

Visualizing the Coupling between Red and Blue Stark States Using Photoionization Microscopy

In nonhydrogenic atoms in a dc electric field, the finite size of the ionic core introduces a coupling between quasibound Stark states that leads to avoided crossings between states that would otherwise cross. Near an avoided crossing, the interacting states may have decay amplitudes that cancel each other, decoupling one of the states from the ionization continuum. This well- known interference narrowing effect, observed as a strongly electric field- dependent decrease in the ionization rate, was previously observed in several atoms. Here we use photoionization microscopy to visualize interference narrowing in helium atoms, thereby explicitly revealing the mechanism by which Stark states decay. The interference narrowing allows measurements of the nodal patterns of red Stark states, which are otherwise not observable due to their intrinsic short lifetime

Particle transport in evolving protoplanetary disks: Implications for results from Stardust

Samples returned from comet 81P/Wild 2 by Stardust confirm that substantial quantities of crystalline silicates were incorporated into the comet at formation. We investigate the constraints that this observation places upon protoplanetary disk physics, assuming that outward transport of particles processed at high temperatures occurs via advection and turbulent diffusion in an evolving disk. We also look for constraints on particle formation locations. Our results are based upon 1D disk models that evolve with time under the action of viscosity and photoevaporation, and track solid transport using an ensemble of individual particle trajectories. We find that two classes of disk model are consistent with the Stardust findings. One class features a high particle diffusivity (a Schmidt number Sc < 1), which suffices to diffuse particles up to 20 microns in size outward against the mean gas flow. For Sc > 1, such models are unlikely to be viable, and significant outward transport requires that the particles of interest settle into a midplane layer that experiences an outward gas flow. In either class of models, the mass of inner disk material that reaches the outer disk is a strong function of the disk's initial compactness. Hence, models of grain transport within steady-state disks underestimate the efficiency of outward transport. Neither model results in sustained outward transport of very large particles exceeding a mm in size. We show that the transport efficiency generally falls off rapidly with time. Hence, high-temperature material must be rapidly incorporated into icy bodies to avoid fallback, and significant radial transport may only occur during the initial phase of rapid disk evolution. It may also vary substantially between disks depending upon their initial mass distributions. We discuss implications for Spitzer observations of crystalline silicates in T Tauri disks.Comment: ApJ, in pres

Solar Wind Abundances of C and O

Quantitative understanding of solar wind (SW) elemental fractionation is required to improve knowledge of the solar nebula abundances from Genesis samples, in particular abundances of volatile elements, depleted in CI chondrites. Ratios of elements with low and high first ionization potential (FIP) in the solar wind, e.g., Fe/He, are higher than photospheric abundances. C, O, and N have intermediate FIP and are thus critical as to whether this fractionation is stepwise or gradual as a function of FIP

Intra-promoter switch of transcription initiation sites in proliferation signaling-dependent RNA metabolism

Global changes in transcriptional regulation and RNA metabolism are crucial features of cancer development. However, little is known about the role of the core promoter in defining transcript identity and post-transcriptional fates, a potentially crucial layer of transcriptional regulation in cancer. In this study, we use CAGE-seq analysis to uncover widespread use of dual-initiation promoters in which non-canonical, first-base-cytosine (C) transcription initiation occurs alongside first-base-purine initiation across 59 human cancers and healthy tissues. C-initiation is often followed by a 5' terminal oligopyrimidine (5'TOP) sequence, dramatically increasing the range of genes potentially subjected to 5'TOP-associated post-transcriptional regulation. We show selective, dynamic switching between purine and C-initiation site usage, indicating transcription initiation-level regulation in cancers. We additionally detail global metabolic changes in C-initiation transcripts that mark differentiation status, proliferative capacity, radiosensitivity, and response to irradiation and to PI3K-Akt-mTOR and DNA damage pathway-targeted radiosensitization therapies in colorectal cancer organoids and cancer cell lines and tissues.</p