On the acceptance, commissioning, and quality assurance of electron FLASH units.

Background & Purpose: FLASH or ultra-high dose rate (UHDR) radiation therapy (RT) has gained attention in recent years for its ability to spare normal tissues relative to conventional dose rate (CDR) RT in various preclinical trials. However, clinical implementation of this promising treatment option has been limited because of the lack of availability of accelerators capable of delivering UHDR RT. Commercial options are finally reaching the market that produce electron beams with average dose rates of up to 1000 Gy/s. We established a framework for the acceptance, commissioning, and periodic quality assurance (QA) of electron FLASH units and present an example of commissioning. Methods: A protocol for acceptance, commissioning, and QA of UHDR linear accelerators was established by combining and adapting standards and professional recommendations for standard linear accelerators based on the experience with UHDR at four clinical centers that use different UHDR devices. Non-standard dosimetric beam parameters considered included pulse width, pulse repetition frequency, dose per pulse, and instantaneous dose rate, together with recommendations on how to acquire these measurements. Results: The 6- and 9-MeV beams of an UHDR electron device were commissioned by using this developed protocol. Measurements were acquired with a combination of ion chambers, beam current transformers (BCTs), and dose-rate-independent passive dosimeters. The unit was calibrated according to the concept of redundant dosimetry using a reference setup. Conclusions: This study provides detailed recommendations for the acceptance testing, commissioning, and routine QA of low-energy electron UHDR linear accelerators. The proposed framework is not limited to any specific unit, making it applicable to all existing eFLASH units in the market. Through practical insights and theoretical discourse, this document establishes a benchmark for the commissioning of UHDR devices for clinical use.

Numerical optimization of longitudinal collimator geometry for novel x-ray field.

Objective. A novel x-ray field produced by an ultrathin conical target is described in the literature. However, the optimal design for an associated collimator remains ambiguous. Current optimization methods using Monte Carlo calculations restrict the efficiency and robustness of the design process. A more generic optimization method that reduces parameter constraints while minimizing computational load is necessary. A numerical method for optimizing the longitudinal collimator hole geometry for a cylindrically-symmetrical x-ray tube is demonstrated and compared to Monte Carlo calculations.Approach. The x-ray phase space was modelled as a four-dimensional histogram differential in photon initial position, final position, and photon energy. The collimator was modeled as a stack of thin washers with varying inner radii. Simulated annealing was employed to optimize this set of inner radii according to various objective functions calculated on the photon flux at a specified plane.Main results. The analytical transport model used for optimization was validated against Monte Carlo calculations using Geant4 via its wrapper, TOPAS. Optimized collimators and the resulting photon flux profiles are presented for three focal spot sizes and five positions of the source. Optimizations were performed with multiple objective functions based on various weightings of precision, intensity, and field flatness metrics. Finally, a select set of these optimized collimators, plus a parallel-hole collimator for comparison, were modeled in TOPAS. The evolution of the radiation field profiles are presented for various positions of the source for each collimator.Significance. This novel optimization strategy proved consistent and robust across the range of x-ray tube settings regardless of the optimization starting point. Common collimator geometries were re-derived using this algorithm while simultaneously optimizing geometry-specific parameters. The advantages of this strategy over iterative Monte Carlo-based techniques, including computational efficiency, radiation source-specificity, and solution flexibility, make it a desirable optimization method for complex irradiation geometries.

Proof-of-concept for a thin conical X-ray target optimized for intensity and directionality for use in a carbon nanotube-based compact X-ray tube.

BACKGROUND: Carbon nanotube-based cold cathode technology has revolutionized the miniaturization of X-ray tubes. However, current applications of these devices required optimization for large, uniform fields with low intensity. PURPOSE: This work investigated the feasibility and radiological characteristics of a novel conical X-ray target optimized for high intensity and high directionality to be used in a compact X-ray tube. METHODS: The proposed device uses an ultrathin, conical tungsten-diamond target that exhibits significant heat loading while maintaining a small focal spot size and promoting forward-directedness of the X-ray field through preferential attenuation of oblique-angled photons. The electrostatic and thermal properties of the theoretical tube were calculated and analyzed using COMSOL Multiphysics software. The production, transport, and calculation of radiological properties associated with the resultant X-ray field were performed using the Geant4 toolkit via its wrapper, TOPAS. RESULTS: Heat transfer analysis of this X-ray tube demonstrated the feasibility of a 200-kV electron beam bombarding the proposed target at a maximum current of 100 mA using a 1-ms symmetric duty cycle. The cathode of the X-ray tube was designed to be segmented into nine switchable electrical segments for modulation of the focal spot size from 0.4- to 10.8-mm. After importing the COMSOL-derived electron beam into TOPAS for X-ray production simulations, radiological analysis of the resultant field demonstrated high levels of intrinsic beam collimation while maintaining high intensity. A maximum dose rate of 17,887 cGy/min was calculated for 1-mm depth in water at 7-cm distance. CONCLUSIONS: The proposed X-ray tube design can create highly directional X-ray fields with superior fluence compared to that of current commercial X-ray tubes of comparable size.

Comprehensive evaluation and new recommendations in the use of Gafchromic EBT3 film.

BACKGROUND: Gafchromic film's unique properties of tissue-equivalence, dose-rate independence, and high spatial resolution make it an attractive choice for many dosimetric applications. However, complicated calibration processes and film handling limits its routine use. PURPOSE: We evaluated the performance of Gafchromic EBT3 film after irradiation under a variety of measurement conditions to identify aspects of film handling and analysis for simplified but robust film dosimetry. METHODS: The short- (from 5 min to 100 h) and long-term (months) film response was evaluated for clinically relevant doses of up to 50 Gy for accuracy in dose determination and relative dose distributions. The dependence of film response on film-read delay, film batch, scanner type, and beam energy was determined. RESULTS: Scanning the film within a 4-h window and using a standard 24-h calibration curve introduced a maximum error of 2% over a dose range of 1-40 Gy, with lower doses showing higher uncertainty in dose determination. Relative dose measurements demonstrated <1 mm difference in electron beam parameters such as depth of 50% of the maximum dose value (R50 ), independent of when the film was scanned after irradiation or the type of calibration curve used (batch-specific or time-specific calibration curve) if the same default scanner was used. Analysis of films exposed over a 5-year period showed that using the red channel led to the lowest variation in the measured net optical density values for different film batches, with doses >10 Gy having the lowest coefficient of variation (<1.7%). Using scanners of similar design produced netOD values within 3% after exposure to doses of 1-40 Gy. CONCLUSIONS: This is the first comprehensive evaluation of the temporal and batch dependence of Gafchromic EBT3 film evaluated on consolidated data over 8 years. The relative dosimetric measurements were insensitive to the type of calibration applied (batch- or time-specific) and in-depth time-dependent dosimetric signal behaviors can be established for film scanned outside of the recommended 16-24 h post-irradiation window. We generated guidelines based on our findings to simplify film handling and analysis and provide tabulated dose- and time-dependent correction factors to achieve this without reducing the accuracy of dose determination.

Dosimetry of a novel focused monoenergetic beam for radiotherapy.

Objective. A novel treatment modality is currently being developed that produces converging monoenergetic x-rays. Conventional application of dosimetric calibration as presented in protocol TG61 is not applicable. Furthermore, the dosimetry of the focal point of the converging beam is on the order of a few millimeters, requiring a high-resolution dosimeter. Here we present a procedure to calibrate radiochromic film for narrow-beam monoenergetic 60 keV photons as well as absolute dosimetry of monoenergetic focused x-rays. A study of the focal spot dose rate after passing through a bone-equivalent material was also done to quantify the effects of heterogeneous materials.Approach.This was accomplished by configuring a polyenergetic beam of equivalent energy using a clinical orthovoltage machine. Calibrated films were then used to perform absolute dosimetry of the converging beam by measuring the beam profile at various depths in water. Main Results.A method for calibrating radiochromic film has been developed and detailed that allows absolute dosimetry of a monoenergetic photon beam. Absolute dosimetry of a focused, mono-energetic beam resulted in a focal spot dose rate of ∼30 cGy min-1at a depth of 5 cm in water.Significance.This work serves to establish a dosimetry protocol for mono-energetic beam absolute dosimetry as well as the use of such a method for measurement of a novel teletherapy modality.

Analysis of a novel X-ray lens for converging beam radiotherapy.

We describe the development and analysis of a new teletherapy modality that, through a novel approach to targeted radiation delivery, has the potential to provide greater conformality than conventional photon-based treatments. The proposed system uses an X-ray lens to reflect photons from a conventional X-ray tube toward a focal spot. The resulting dose distributions have a highly localized peak dose, with lower doses in the converging radiation cone. Physical principles governing the design of this system are presented, along with a series of measurements analyzing various characteristics of the converging beam. The beam was designed to be nearly monoenergetic (~ 59 keV), with an energy bandwidth of approximately 10 keV allowing for treatment energies lower than conventional therapies. The focal spot was measured to be approximately 2.5 cm long and 4 mm wide. Mounting the proposed X-ray delivery system on a robotic arm would allow sub-millimeter accuracy in focal spot positioning, resulting in highly conformal dose distribution via the optimal placement of individual focal spots within the target volume. Aspects of this novel radiation beam are discussed considering their possible clinical application as a treatment approach that takes maximum advantage of the unique properties afforded by converging X-ray beam therapy.

Postpubertal spermatogonial stem cell transplantation restores functional sperm production in rhesus monkeys irradiated before and after puberty.

BACKGROUND: Cancer treatment of prepubertal patients impacts future fertility due to the abolition of spermatogonial stem cells (SSCs). In macaques, spermatogenesis could be regenerated by intratesticular transplantation of SSCs, but no studies have involved cytotoxic treatment before puberty and transplantation after puberty, which would be the most likely clinical scenario. OBJECTIVES: To evaluate donor-derived functional sperm production after SSC transplantation to adult monkeys that had received testicular irradiation during the prepubertal period. MATERIALS AND METHODS: We obtained prepubertal testis tissue by unilaterally castrating six prepubertal monkeys and 2 weeks later irradiated the remaining testes with 6.9 Gy. However, because spermatogenic recovery was observed, we irradiated them again 14 months later with 7 Gy. Three of the monkeys were treated with GnRH-antagonist (GnRH-ant) for 8 weeks. The cryopreserved testis cells from the castrated testes were then allogeneically transplanted into the intact testes of all monkeys. Tissues were harvested 10 months later for analyses. RESULTS: In three of the six monkeys, 61%, 38%, and 11% of the epididymal sperm DNA were of the donor genotype. The ability to recover donor-derived sperm production was not enhanced by the GnRH-ant pretreatment. However, the extent of filling seminiferous tubules during the transplantation procedure was correlated with the eventual production of donor spermatozoa. The donor epididymal spermatozoa from the recipient with 61% donor contribution were capable of fertilizing rhesus eggs and forming embryos. Although the transplantation was done into the rete testis, two GnRH-ant-treated monkeys, which did not produce donor-derived epididymal spermatozoa, displayed irregular tubular cords in the interstitium containing testicular spermatozoa derived from the transplanted donor cells. DISCUSSION AND CONCLUSION: The results further support that sperm production can be restored in non-human primates from tissues cryopreserved prior to prepubertal and post-pubertal gonadotoxic treatment by transplantation of these testicular cells after puberty into seminiferous tubules.

Devices for dosimetric measurements and quality assurance of the Xstrahl 300 orthovoltage unit.

The Xstrahl 300 orthovoltage unit is designed to deliver kilovoltage radiation therapy using the appositional technique. However, it is not equipped with some typical linear accelerator features, such as mechanical distance indicator and crosshair projection, which are useful for facilitating equipment setup during various quality assurance (QA) and research activities. Therefore, we designed and constructed slip-in devices to facilitate QA for dosimetric measurements of our Xstrahl 300 unit. These include: (a) an ion chamber positioning system for dosimetric measurements, (b) a mechanical pointer for setting dosimeter distance to a nominal 50 cm, and (c) a crosshair projector with built-in light to facilitate alignment of dosimeter to the center of the radiation field. These devices provide a high degree of setup reproducibility thereby minimizing setup errors. We used these devices to perform QA of the Xstrahl 300 orthovoltage unit. One of the QA tests we perform is a constancy check of beam output and energy. Our data since start of clinical use of this unit (approximately 2.5 yr) show dose outputs to be remarkably reproducible (2σ = ±0.4%) for all three clinical beams (75, 125, and 250 kVp). These devices have provided both convenience and high-precision during the unit's commissioning, and continue to provide the same for various QA activities on the Xstrahl 300 orthovoltage unit.

Author Correction: Ultra high dose rate (35 Gy/sec) radiation does not spare the normal tissue in cardiac and splenic models of lymphopenia and gastrointestinal syndrome.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Restoration of functional sperm production in irradiated pubertal rhesus monkeys by spermatogonial stem cell transplantation.

BACKGROUND: In male pre-pubertal cancer patients, radiation and chemotherapy impact future fertility by eradication of spermatogonial stem cells (SSCs). In macaques, spermatogenesis could be regenerated by intratesticular transplantation of SSCs, but only a small percentage of spermatozoa produced were of donor origin. Transient hormone suppression with a GnRH antagonist (GnRH-ant) enhanced spermatogenic recovery from transplanted SSCs. OBJECTIVES: To evaluate donor-derived and endogenous spermatogenic recovery after SSC transplantation into irradiated monkeys and to test whether hormone suppression around the time of transplantation facilitates spermatogenic recovery. MATERIALS AND METHODS: Testes of 15 adult rhesus monkeys were irradiated with 7 Gy and 4 months later transplanted, to one of the testes, with cryopreserved testicular cells containing SSCs from unrelated monkeys. Monkeys were either treated with GnRH-ant for 8 weeks before transplantation, GnRH-ant from 4 weeks before to 4 weeks after transplantation, or with no GnRH-ant. Tissues were harvested 10 months after transplantation. RESULTS: Two of the 15 monkeys, a control and a pre-transplantation GnRH-ant-treated, showed substantially higher levels of testicular spermatogenesis and epididymal sperm output in the transplanted side as compared to the untransplanted. Over 84% of epididymal spermatozoa on the transplanted side had the donor genotype and were capable of fertilizing eggs after intracytoplasmic sperm injection forming morulae of the donor paternal origin. Low levels of donor spermatozoa (~1%) were also identified in the epididymis of three additional monkeys. Transplantation also appeared to enhance endogenous spermatogenesis. DISCUSSION AND CONCLUSION: We confirmed that SSC transplantation can be used for restoration of fertility in male cancer survivors exposed to irradiation as a therapeutic agent. The success rate of this procedure, however, is low. The success of filling the tubules with the cell suspension, but not the GnRH-ant treatment, was related to the level of colonization by transplanted cells.

Effect of Radiation on DCE-MRI Pharmacokinetic Parameters in a Rabbit Model of Compromised Maxillofacial Wound Healing: A Pilot Study.

PURPOSE: Osteoradionecrosis (ORN), a potentially debilitating complication of maxillofacial radiation, continues to present a challenging clinical scenario, with limited treatment options that often fail. Translational animal models that can accurately mimic the human characteristics of the condition are lacking. In the present pilot study, we aimed to characterize the effects of radiation on the dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) pharmacokinetic parameters in a rabbit model of compromised maxillofacial wound healing to determine its potential as a translational model of ORN. MATERIALS AND METHODS: An experimental group underwent fractionated radiation of the mandible totaling 36 Gy. At 4 weeks after irradiation, the experimental and control groups (n = 8 rabbits each) underwent a surgical procedure to create a critical size defect in the mandibular bone. DCE-MRI scans were acquired 1 week after arrival (baseline; time point 1), 4 weeks after completion of irradiation in the experimental group (just before surgery, time point 2), and 4 weeks after surgery (time point 3). RESULTS: No differences in the analyzed DCE-MRI parameters were noted within the experimental or control group between the baseline values (time point 1) and those after irradiation (time point 2). The whole blood volume fraction (vb) in the experimental group was increased compared with that in the control group after irradiation (time point 2; P < .05). After surgery (time point 3), both the forward flux rate of contrast from blood plasma and the extracellular extravascular space and the vb were increased in the control group compared with the experimental group (P < .05). CONCLUSIONS: The results of the present study suggest that DCE-MRI of a rabbit model of compromised maxillofacial wound healing could reflect the DCE-MRI characteristics of human patients with ORN and those at risk of developing the condition. Future studies will focus on further characterization of this rabbit model as a translational preclinical model of ORN.

A Mail Audit Independent Peer Review System for Dosimetry Verification of a Small Animal Irradiator.

Dedicated precision orthovoltage small animal irradiators have become widely available in the past decade and are commonly used for radiation biology research. However, there is a lack of dosimetric standardization among these irradiators, which affects the reproducibility of radiation-based animal studies. The purpose of this study was to develop a mail-based, independent peer review system to verify dose delivery among institutions using X-RAD 225Cx irradiators (Precision X-Ray, North Branford, CT). A robust, user-friendly mouse phantom was constructed from high-impact polystyrene and designed with dimensions similar to those of a typical laboratory mouse. The phantom accommodates three thermoluminescent dosimeters (TLDs) to measure dose. The mouse peer review system was commissioned in a small animal irradiator using anterior-posterior and posterior-anterior beams of 225 kVp and then mailed to three institutions to test the feasibility of the audit service. The energy correction factor for TLDs in the mouse phantom was derived to validate the delivered dose using this particular animal irradiation system. This feasibility study indicated that three institutions were able to deliver a radiation dose to the mouse phantom within ±10% of the target dose. The developed mail audit independent peer review system for the verification of mouse dosimetry can be expanded to characterize other commercially available orthovoltage irradiators, thereby enhancing the reproducibility of studies employing these irradiators.

A novel patient-derived orthotopic xenograft model of esophageal adenocarcinoma provides a platform for translational discoveries.

Mouse models of gastroesophageal junction (GEJ) cancer strive to recapitulate the intratumoral heterogeneity and cellular crosstalk within patient tumors to improve clinical translation. GEJ cancers remain a therapeutic challenge due to the lack of a reliable mouse model for preclinical drug testing. In this study, a novel patient-derived orthotopic xenograft (PDOX) was established from GEJ cancer via transabdominal surgical implantation. Patient tumor was compared to subcutaneously implanted patient-derived tumor xenograft (PDX) and PDOX by Hematoxylin and Eosin staining, immunohistochemistry and next-generation sequencing. Treatment efficacy studies of radiotherapy were performed. We observed that mechanical abrasion of mouse GEJ prior to surgical implantation of a patient-derived tumor in situ promotes tumor engraftment (100%, n=6). Complete PDOX engraftment was observed with rapid intra- and extraluminal tumor growth, as evidenced by magnetic resonance imaging. PDOXs contain fibroblasts, tumor-associated macrophages, immune and inflammatory cells, vascular and lymphatic vessels. Stromal hallmarks of aggressive GEJ cancers are recapitulated in a GEJ PDOX mouse model. PDOXs demonstrate tumor invasion into vasculature and perineural space. Next-generation sequencing revealed loss of heterozygosity with very high allelic frequency in NOTCH3, TGFB1, EZH2 and KMT2C in the patient tumor, the subcutaneous PDX and the PDOX. Immunohistochemical analysis of Her2/neu (also known as ERBB2), p53 (also known as TP53) and p16 (also known as CDKN2A) in PDX and PDOX revealed maintenance of expression of proteins found in patient tumors, but membranous EGFR overexpression in patient tumor cells was absent in both xenografts. Targeted radiotherapy in this model suggested a decrease in size by 61% according to Response Evaluation Criteria in Solid Tumors (RECIST), indicating a partial response to radiation therapy. Our GEJ PDOX model exhibits remarkable fidelity to human disease and captures the precise tissue microenvironment present within the local GEJ architecture, providing a novel tool for translating findings from studies on human GEJ cancer. This model can be applied to study metastatic progression and to develop novel therapeutic approaches for the treatment of GEJ cancer.This article has an associated First Person interview with the first author of the paper.

Ultra high dose rate (35 Gy/sec) radiation does not spare the normal tissue in cardiac and splenic models of lymphopenia and gastrointestinal syndrome.

Recent reports have shown that very high dose rate radiation (35-100 Gy/second) referred to as FLASH tends to spare the normal tissues while retaining the therapeutic effect on tumor. We undertook a series of experiments to assess if ultra-high dose rate of 35 Gy/second can spare the immune system in models of radiation induced lymphopenia. We compared the tumoricidal potency of ultra-high dose rate and conventional dose rate radiation using a classical clonogenic assay in murine pancreatic cancer cell lines. We also assessed the lymphocyte sparing potential in cardiac and splenic irradiation models of lymphopenia and assessed the severity of radiation-induced gastrointestinal toxicity triggered by the two dose rate regimes in vivo. Ultra-high dose rate irradiation more potently induces clonogenic cell death than conventional dose rate irradiation with a dose enhancement factor at 10% survival (DEF10) of 1.310 and 1.365 for KPC and Panc02 cell lines, respectively. Ultra-high dose rate was equally potent in depleting CD3, CD4, CD8, and CD19 lymphocyte populations in both cardiac and splenic irradiation models of lymphopenia. Radiation-induced gastrointestinal toxicity was more pronounced and mouse survival (7 days vs. 15 days, p = 0.0001) was inferior in the ultra-high dose rate arm compared to conventional dose rate arm. These results suggest that, contrary to published data in other models of radiation-induced acute and chronic toxicity, dose rates of 35 Gy/s do not protect mice from the detrimental side effects of irradiation in our models of cardiac and splenic radiation-induced lymphopenia or gastrointestinal mucosal injury.

Exploiting Arginine Auxotrophy with Pegylated Arginine Deiminase (ADI-PEG20) to Sensitize Pancreatic Cancer to Radiotherapy via Metabolic Dysregulation.

Distinct metabolic vulnerabilities of cancer cells compared with normal cells can potentially be exploited for therapeutic targeting. Deficiency of argininosuccinate synthetase-1 (ASS1) in pancreatic cancers creates auxotrophy for the semiessential amino acid arginine. We explored the therapeutic potential of depleting exogenous arginine via pegylated arginine deiminase (ADI-PEG20) treatment as an adjunct to radiotherapy. We evaluated the efficacy of treatment of human pancreatic cancer cell lines and xenografts with ADI-PEG20 and radiation via clonogenic assays and tumor growth delay experiments. We also investigated potential mechanisms of action using reverse-phase protein array, Western blotting, and IHC and immunofluorescence staining. ADI-PEG20 potently radiosensitized ASS1-deficient pancreatic cancer cells (MiaPaCa-2, Panc-1, AsPc-1, HPAC, and CaPan-1), but not ASS1-expressing cell lines (Bxpc3, L3.6pl, and SW1990). Reverse phase protein array studies confirmed increased expression of proteins related to endoplasmic reticulum (ER) stress and apoptosis, which were confirmed by Western blot analysis. Inhibition of ER stress signaling with 4-phenylbutyrate abrogated the expression of ER stress proteins and reversed radiosensitization by ADI-PEG20. Independent in vivo studies in two xenograft models confirmed significant tumor growth delays, which were associated with enhanced expression of ER stress proteins and apoptosis markers and reduced expression of proliferation and angiogenesis markers. ADI-PEG20 augmented the effects of radiation by triggering the ER stress pathway, leading to apoptosis in pancreatic tumor cells.

Targeting cyclin-dependent kinase 9 by a novel inhibitor enhances radiosensitization and identifies Axl as a novel downstream target in esophageal adenocarcinoma.

Cyclin-dependent kinase 9 (CDK9) transcriptionally regulates several proteins and cellular pathways central to radiation induced tissue injury. We investigated a role of BAY1143572, a new highly specific CDK9 inhibitor, as a sensitizer to radiation in esophageal adenocarcinoma. In vitro synergy between the CDK9 inhibitor and radiation was evaluated by clonogenic assay. In vivo synergy between the CDK9 inhibitor and radiation was assessed in multiple xenograft models including a patient's tumor derived xenograft (PDX). Reverse phase protein array (RPPA), western blotting, immunohistochemistry, and qPCR were utilized to identify and validate targets of the CDK9 inhibitor. The CDK9 inhibitor plus radiation significantly reduced growth of FLO-1, SKGT4, OE33, and radiation resistant OE33R xenografts and PDXs as compared to the cohorts treated with either single agent CDK9 inhibitor or radiation alone. RPPA identified Axl as a candidate target of CDK9 inhibition. Western blot and qPCR demonstrated reduced Axl mRNA (p = 0.02) and protein levels after treatment with CDK9 inhibitor with or without radiation in FLO-1 and SKGT4 cells. Axl protein expression in FLO-1 xenografts treated with combination of CDK9 inhibitor and radiation was significantly lower than the xenografts treated with radiation alone (p = 0.003). Clonogenic assay performed after overexpression of Axl in FLO-1 and SKGT4 cells enhanced radiosensitization by the CDK9 inhibitor, suggesting dependency of radiosensitization effects of the CDK9 inhibitor on Axl. In conclusion, these findings indicate that targeting CDK9 by BAY1143572 significantly enhances the effects of radiation and Axl is a novel downstream target of CDK9 in esophageal adenocarcinoma.

Transient enhancement of p53 activity protects from radiation-induced gastrointestinal toxicity.

Gastrointestinal (GI) syndrome is a serious side effect and dose-limiting toxicity observed in patients undergoing lower-abdominal radiotherapy. Previous mouse studies show that p53 gene dosage determines susceptibility to GI syndrome development. However, the translational relevance of p53 activity has not been addressed. Here, we used a knock-in mouse in which the p53-Mdm2 negative feedback loop is genetically disrupted. These mice retain biallelic p53 and thus, normal basal p53 levels and activity. However, due to the lack of p53-mediated Mdm2 transcription, irradiated Mdm2P2/P2 mice exhibit enhanced acute p53 activity, which protects them from GI failure. Intestinal crypt cells residing in the +4 and higher positions exhibit decreased apoptosis, increased p21 expression, and hyperproliferation to reinstate intestinal integrity. Correspondingly, pharmacological augmentation of p53 activity in wild-type mice with an Mdm2 inhibitor protects against GI toxicity without affecting therapeutic outcome. Our results suggest that transient disruption of the p53-Mdm2 interaction to enhance p53 activity could be a viable prophylactic strategy for alleviating GI syndrome in patients undergoing radiotherapy.

Selective EGLN Inhibition Enables Ablative Radiotherapy and Improves Survival in Unresectable Pancreatic Cancer.

When pancreatic cancer cannot be removed surgically, patients frequently experience morbidity and death from progression of their primary tumor. Radiation therapy (RT) cannot yet substitute for an operation because radiation causes fatal bleeding and ulceration of the nearby stomach and intestines before achieving tumor control. There are no FDA-approved medications that prevent or reduce radiation-induced gastrointestinal injury. Here, we overcome this fundamental problem of anatomy and biology with the use of the oral EGLN inhibitor FG-4592, which selectively protects the intestinal tract from radiation toxicity without protecting tumors. A total of 70 KPC mice with autochthonous pancreatic tumors received oral FG-4592 or vehicle control ± ablative RT to a cumulative 75 Gy administered in 15 daily fractions to a limited tumor field. Although ablative RT reduced complications from local tumor progression, fatal gastrointestinal bleeding was observed in 56% of mice that received high-dose RT with vehicle control. However, radiation-induced bleeding was completely ameliorated in mice that received high-dose RT with FG-4592 (0% bleeding, P < 0.0001 compared with vehicle). Furthermore, FG-4592 reduced epithelial apoptosis by half (P = 0.002) and increased intestinal microvessel density by 80% compared with vehicle controls. EGLN inhibition did not stimulate cancer growth, as treatment with FG-4592 alone, or overexpression of HIF2 within KPC tumors independently improved survival. Thus, we provide a proof of concept for the selective protection of the intestinal tract by the EGLN inhibition to enable ablative doses of cytotoxic therapy in unresectable pancreatic cancer by reducing untoward morbidity and death from radiation-induced gastrointestinal bleeding. SIGNIFICANCE: Selective protection of the intestinal tract by EGLN inhibition enables potentially definitive doses of radiation therapy. This might allow radiation to be a surgical surrogate for unresectable pancreatic cancer.Graphical Abstract: http://cancerres.aacrjournals.org/content/canres/79/9/2327/F1.large.jpg.

Merging Orthovoltage X-Ray Minibeams spare the proximal tissues while producing a solid beam at the target.

Conventional radiation therapy of brain tumors often produces cognitive deficits, particularly in children. We investigated the potential efficacy of merging Orthovoltage X-ray Minibeams (OXM). It segments the beam into an array of parallel, thin (~0.3 mm), planar beams, called minibeams, which are known from synchrotron x-ray experiments to spare tissues. Furthermore, the slight divergence of the OXM array make the individual minibeams gradually broaden, thus merging with their neighbors at a given tissue depth to produce a solid beam. In this way the proximal tissues, including the cerebral cortex, can be spared. Here we present experimental results with radiochromic films to characterize the method's dosimetry. Furthermore, we present our Monte Carlo simulation results for physical absorbed dose, and a first-order biologic model to predict tissue tolerance. In particular, a 220-kVp orthovoltage beam provides a 5-fold sharper lateral penumbra than a 6-MV x-ray beam. The method can be implemented in arc-scan, which may include volumetric-modulated arc therapy (VMAT). Finally, OXM's low beam energy makes it ideal for tumor-dose enhancement with contrast agents such as iodine or gold nanoparticles, and its low cost, portability, and small room-shielding requirements make it ideal for use in the low-and-middle-income countries.

Development and Characterization of a Rabbit Model of Compromised Maxillofacial Wound Healing.

IMPACT STATEMENT: Maxillofacial defects often present the clinical challenge of a compromised wound bed. Preclinical evaluation of tissue engineering techniques developed to facilitate healing and reconstruction typically involves animal models with ideal wound beds. The healthy wound bed scenario does not fully mimic the complex clinical environment in patients, which can lead to technology failure when translating from preclinical in vivo research to clinical use. The reported preclinical animal model of compromised wound healing enables investigation of tissue engineering technologies in a more clinically relevant scenario, potentially fostering translation of promising results in preclinical research to patients.