SGK1 signaling promotes glucose metabolism and survival in extracellular matrix detached cells.

Loss of integrin-mediated attachment to extracellular matrix (ECM) proteins can trigger a variety of cellular changes that affect cell viability. Foremost among these is the activation of anoikis, caspase-mediated cell death induced by ECM detachment. In addition, loss of ECM attachment causes profound alterations in cellular metabolism, which can lead to anoikis-independent cell death. Here, we describe a surprising role for serum and glucocorticoid kinase-1 (SGK1) in the promotion of energy production when cells are detached. Our data demonstrate that SGK1 activation is necessary and sufficient for ATP generation during ECM detachment and anchorage-independent growth. More specifically, SGK1 promotes a substantial elevation in glucose uptake because of elevated GLUT1 transcription. In addition, carbon flux into the pentose phosphate pathway (PPP) is necessary to accommodate elevated glucose uptake and PPP-mediated glyceraldehyde-3-phosphate (G3P) is necessary for ATP production. Thus, our data show SGK1 as master regulator of glucose metabolism and cell survival during ECM-detached conditions.

Dosimetric characterization of the iBEAM evo carbon fiber couch for radiotherapy.

PURPOSE: This study characterizes the dosimetric properties of the iBEAM evo carbon fiber couch manufactured by Medical Intelligence and examines the accuracy of the CMS XiO and Nucletron Oncentra Masterplan (OMP) treatment planning systems for calculating beam attenuation due to the presence of the couch. METHODS: To assess the homogeneity of the couch, it was CT scanned at isocentric height and a number of signal intensity profiles were generated and analyzed. To simplify experimental procedures, surface dose and central axis depth dose measurements were performed in a solid water slab phantom using Gafchromic film for 6 and 10 MV photon beams at gantry angles of 0 degree (normal incidence), 30 degrees, and 60 degrees with an inverted iBEAM couch placed on top of the phantom. Attenuation measurements were performed in a cylindrical solid water phantom with an ionization chamber positioned at the isocenter. Measurements were taken for gantry angles from 0 degree to 90 degrees in 10 degrees increments for both 6 and 10 MV photon beams. This setup was replicated in the XiO and OMP treatment planning systems. Dose was calculated using the pencil beam, collapsed cone, convolution, and superposition algorithms. RESULTS: The CT scan of the couch showed that it was uniformly constructed. Surface dose increased by (510 +/-0)% for a 6 MV beam and (600 +/- 20)% for a 10 MV beam passing through the couch at normal incidence. Obliquely incident beams resulted in a higher surface dose compared to normally incident beams for both open fields and fields with the couch present. Depth dose curves showed that the presence of the couch resulted in an increase in dose in the build up region. For 6 and 10 MV beams incident at 60 degrees, nearly all skin sparing was lost. Attenuation measurements derived using the ionization chamber varied from 2.7% (0 degree) to a maximum of 4.6% (50 degrees) for a 6 MV beam and from 1.9% (0 degree) to a maximum of 4.0% (50 degrees) for a 10 MV beam. The pencil beam and convolution algorithms failed to accurately calculate couch attenuation. The collapsed cone and superposition algorithms calculated attenuation within an absolute error of +/- 1.2% for 6 MV and +/- 0.8% for 10 MV for gantry angles from 0 degree to 40 degrees. Some differences in attenuation were observed dependent on how the couch was contoured. CONCLUSIONS: These results demonstrate that the presence of the iBEAM evo carbon fiber couch increases the surface dose and dose in the build up region. The inclusion of the couch in the planning scan is limited by the field of view employed and the couch height at the time of CT scanning.

UK audit of target volume and organ at risk delineation and dose optimisation for cervix radiotherapy treatments.

OBJECTIVE: Assessment of the extent of variation in delineations and dose optimisation performed at multiple UK centres as a result of interobserver variation and protocol differences. METHODS: CT/MR images of 2 cervical cancer patients previously treated with external beam radiotherapy (EBRT) and Brachytherapy were distributed to 11 UK centres. Centres delineated structures and produced treatment plans following their local protocol. Organ at risk delineations were assessed dosimetrically through application of the original treatment plan and target volume delineations were assessed in terms of variation in absolute volume and length, width and height. Treatment plan variation was assessed across all centres and across centres that followed EMBRACE II. Treatment plans were assessed using total EQD2 delivered and were compared to EMBRACE II dose aims. Variation in combined intracavitary/interstitial brachytherapy treatments was also assessed. RESULTS: Brachytherapy target volume delineations contained variation due to differences in protocol used, window/level technique and differences in interpretations of grey zones. Planning target volume delineations were varied due to protocol differences and extended parametrial tissue inclusion. All centres met EMBRACE II plan aims for PTV V95 and high-riskclinical target volume D90 EQD2, despite variation in prescription dose, fractionation and treatment technique. CONCLUSION: Brachytherapy target volume delineations are varied due to differences in contouring guidelines and protocols used. Planning target volume delineations are varied due to the uncertainties surrounding the extent of parametrial involvement. Dosimetric optimisation is sufficient across all centres to satisfy EMBRACE II planning aims despite significant variation in protocols used. ADVANCES IN KNOWLEDGE: Previous multi-institutional audits of cervical cancer radiotherapy practices have been performed in Europe and the USA. This study is the first of its kind to be performed in the UK.

Ten-year longitudinal health-related quality of life following iodine-125 brachytherapy monotherapy for localized prostate cancer.

PURPOSE: This prospective longitudinal study quantifies health-related quality of life (HRQoL) up to 10 years following permanent iodine-125 (125I) prostate brachytherapy alone for localized prostate cancer. MATERIAL AND METHODS: In total, 120 patients completed a validated expanded prostate cancer index composite (EPIC) questionnaire pre-treatment and at 8 time points after treatment (6 weeks, 6, 10, 18 months, and 2, 3, 5, 10 years). At each time point, clinically relevant small, moderate, and severe declines in HRQoL were defined as 0.2-0.5 SD, 0.5-0.8 SD, and > 0.8 SD of baseline function for each of urinary, bowel, and sexual domains, respectively. RESULTS: Response rates in the first two years were > 90%, but thereafter dropped to 75% and 48% at 5 and 10 years, respectively. 50 patients (41.6%) responded at all stages. Maximal deterioration in mean urinary and sexual summary scores was noted 6 weeks after implant, with severe urinary symptoms and moderate bowel/sexual symptoms. At 6 months, urinary and bowel quality of life (QoL) had improved to mild impairment, which then fully resolved at 10 months. Sexual QoL remained mildly impaired throughout the 10 years of follow-up. At 10 years, new mild impairment of urinary and bowel QoL was found. CONCLUSIONS: Clinically mild changes in urinary, bowel, and sexual QoL are found 10 years after 125I monotherapy. The impairment in sexual function persists from treatment, but urinary and bowel symptoms are new at 10 years.

A comparison of outcomes for patients with intermediate and high risk prostate cancer treated with low dose rate and high dose rate brachytherapy in combination with external beam radiotherapy.

INTRODUCTION: There is evidence to support use of external beam radiotherapy (EBRT) in combination with both low dose rate brachytherapy (LDR-EBRT) and high dose rate brachytherapy (HDR-EBRT) to treat intermediate and high risk prostate cancer. METHODS: Men with intermediate and high risk prostate cancer treated using LDR-EBRT (treated between 1996 and 2007) and HDR-EBRT (treated between 2007 and 2012) were identified from an institutional database. Multivariable analysis was performed to evaluate the relationship between patient, disease and treatment factors with biochemical progression free survival (bPFS). RESULTS: 116 men were treated with LDR-EBRT and 171 were treated with HDR-EBRT. At 5 years, bPFS was estimated to be 90.5% for the LDR-EBRT cohort and 77.6% for the HDR-EBRT cohort. On multivariable analysis, patients treated with HDR-EBRT were more than twice as likely to experience biochemical progression compared with LDR-EBRT (HR 2.33, 95% CI 1.12-4.07). Patients with Gleason ≥8 disease were more than five times more likely to experience biochemical progression compared with Gleason 6 disease (HR 5.47, 95% CI 1.26-23.64). Cumulative incidence of ≥grade 3 genitourinary and gastrointestinal toxicities for the LDR-EBRT and HDR-EBRT cohorts were 8% versus 4% and 5% versus 1% respectively, although these differences did not reach statistical significance. CONCLUSION: LDR-EBRT may provide more effective PSA control at 5 years compared with HDR-EBRT. Direct comparison of these treatments through randomised trials are recommended to investigate this hypothesis further.

Efficacy and toxicity outcomes for patients treated with focal salvage high dose rate brachytherapy for locally recurrent prostate cancer.

INTRODUCTION: Isolated local recurrence of prostate cancer following primary radiotherapy or brachytherapy may be treated with focal salvage high dose rate brachytherapy, although there remains an absence of high quality evidence to support this approach. METHODS: Men with prostate cancer treated consecutively between 2015 and 2018 using 19 Gy in a single fraction high dose rate brachytherapy (HDR) for locally recurrent prostate cancer were identified from an institutional database. Univariable analysis was performed to evaluate the relationship between patient, disease and treatment factors with biochemical progression free survival (bPFS). RESULTS: 43 patients were eligible for evaluation. Median follow up duration was 26 months (range 1-60). Median bPFS was 35 months (95% confidence interval 25.6-44.4). Kaplan-Meier estimates for bPFS at 1, 2 and 3 years post salvage were 95.2%, 70.6% and 41.8% respectively. On univariable Cox regression analysis, only nadir PSA was significantly associated with bPFS although the majority of patients were also treated with androgen deprivation therapy. Only one late grade 3 genitourinary toxicity was observed. CONCLUSION: Focal salvage HDR brachytherapy may provide good biochemical control with a low risk of severe toxicity. Further evaluation within clinical trials are needed to establish its role in the management of locally recurrent prostate cancer.

Ras-ling with new therapeutic targets for metastasis.

Successful cancer metastasis relies on the ability of cancer cells to survive independently of attachment to the extracellular matrix (ECM) and to overcome ECM-detachment-induced death programs. This can be accomplished through activating mutations in cellular oncogenes that subsequently lead to the inhibition of anoikis and to alterations in productive metabolism. One example of such an oncogene is Ras which is found to be mutated and hyperactivated in a variety of distinct cancers. Despite numerous studies on Ras, the precise molecular mechanisms that facilitate survival during ECM-detachment remain poorly understood. Recently, we discovered that ECM-detached cells harboring oncogenic Ras mutations require signaling through the PI(3)K/SGK1 signaling axis to promote survival. Furthermore, we found that oncogenic Ras can concurrently diminish PHLPP1 phosphatase levels, which results in a decrease in p38 MAPK-mediated activation of anoikis. Thus, our data suggest that cancer cells with activating Ras mutations can survive during ECM-detachment using downstream effector molecules that modulate distinct pathways. Overall, these data suggest that new therapeutic interventions that aim to mitigate SGK1 signaling and activate the p38 MAPK activity may aid in specifically targeting and eliminating metastatic cancer cells.

RIPK1-mediated induction of mitophagy compromises the viability of extracellular-matrix-detached cells.

For cancer cells to survive during extracellular matrix (ECM) detachment, they must inhibit anoikis and rectify metabolic deficiencies that cause non-apoptotic cell death. Previous studies in ECM-detached cells have linked non-apoptotic cell death to reactive oxygen species (ROS) generation, although the mechanistic underpinnings of this link remain poorly defined. Here, we uncover a role for receptor-interacting protein kinase 1 (RIPK1) in the modulation of ROS and cell viability during ECM detachment. We find that RIPK1 activation during ECM detachment results in mitophagy induction through a mechanism dependent on the mitochondrial phosphatase PGAM5. As a consequence of mitophagy, ECM-detached cells experience diminished NADPH production in the mitochondria, and the subsequent elevation in ROS levels leads to non-apoptotic death. Furthermore, we find that antagonizing RIPK1/PGAM5 enhances tumour formation in vivo. Thus, RIPK1-mediated induction of mitophagy may be an efficacious target for therapeutics aimed at eliminating ECM-detached cancer cells.

Metabolism during ECM Detachment: Achilles Heel of Cancer Cells?

Integrin-mediated attachment to the extracellular matrix (ECM) is required to combat the induction of programmed cell death in a variety of distinct cell types. If cells fail to maintain proper ECM attachment, they become subject to elimination via an apoptotic cell death program known as anoikis. However, anoikis inhibition is not sufficient to promote the long-term survival of ECM-detached cells. Several recent studies have unveiled the profound (anoikis-independent) impact of cell metabolism on the viability of ECM-detached cells. Thus, we posit that, during metastatic dissemination (when cancer cells are exposed to periods of ECM detachment), cancer cells must alter their metabolism in a fashion that promotes survival and ultimately contributes to metastatic outgrowth.

Modelling second malignancy risks from low dose rate and high dose rate brachytherapy as monotherapy for localised prostate cancer.

BACKGROUND AND PURPOSE: To estimate the risks of radiation-induced rectal and bladder cancers following low dose rate (LDR) and high dose rate (HDR) brachytherapy as monotherapy for localised prostate cancer and compare to external beam radiotherapy techniques. MATERIALS AND METHODS: LDR and HDR brachytherapy monotherapy plans were generated for three prostate CT datasets. Second cancer risks were assessed using Schneider's concept of organ equivalent dose. LDR risks were assessed according to a mechanistic model and a bell-shaped model. HDR risks were assessed according to a bell-shaped model. Relative risks and excess absolute risks were estimated and compared to external beam techniques. RESULTS: Excess absolute risks of second rectal or bladder cancer were low for both LDR (irrespective of the model used for calculation) and HDR techniques. Average excess absolute risks of rectal cancer for LDR brachytherapy according to the mechanistic model were 0.71 per 10,000 person-years (PY) and 0.84 per 10,000 PY respectively, and according to the bell-shaped model, were 0.47 and 0.78 per 10,000 PY respectively. For HDR, the average excess absolute risks for second rectal and bladder cancers were 0.74 and 1.62 per 10,000 PY respectively. The absolute differences between techniques were very low and clinically irrelevant. Compared to external beam prostate radiotherapy techniques, LDR and HDR brachytherapy resulted in the lowest risks of second rectal and bladder cancer. CONCLUSIONS: This study shows both LDR and HDR brachytherapy monotherapy result in low estimated risks of radiation-induced rectal and bladder cancer. LDR resulted in lower bladder cancer risks than HDR, and lower or similar risks of rectal cancer. In absolute terms these differences between techniques were very small. Compared to external beam techniques, second rectal and bladder cancer risks were lowest for brachytherapy.