BACKGROUND:Orchialgia is a rare but significant complication experienced after open and minimally invasive inguinal hernia repairs with or without mesh. Causes have been theorized to include trauma to the spermatic cord, mesh-induced inflammation, vascular insult, fibrosis or mesh scarring of the cord, and neuropathic injury to the autonomic paravasal nerve fibers supplying the testicle. Though less frequently described than inguinodynia, orchialgia can be severely debilitating. METHODS:Retrospective review of consecutive adult patients who underwent surgical intervention for inguinodynia with coexisting orchialgia between September 2011 and April 2023. Primary outcome was percent improvement of orchialgia, reported by patients in postoperative follow up. Secondary outcomes included surgical approach, type of neurectomy performed, the need for orchiectomy, and remedial operations. RESULTS:A total of 173 patients were identified. Of this cohort, 89.6% (155) underwent paravasal neurectomy, 96% (166) underwent inguinal neurectomy, 14% (24) underwent orchiectomy, and 5.2% (9) underwent vasectomy. These operations were conducted via an open approach in 18% (31) and via hybrid approach in 62% (108). Improvement was noted in 99% (173) of cases, with greater than 50% improvement in pain in 64% (111). CONCLUSION:Orchialgia is less common than inguinodynia but is encountered in approximately 10% of patients with chronic pain after inguinal hernia repair resulting from all variants of operative repairs. Utilization of a patient-tailored surgical approach to address the likely mechanisms of testicular pain after prior inguinal hernia repair allows for favorable surgical results. A careful examination and assessment of the potential mechanisms of orchialgia after inguinal hernia repair allows for identification of specific causes and potential options for remediation.

Supplementary Figures S1-S2 from Somatic Mutations in the Tyrosine Kinase Domain of Epidermal Growth Factor Receptor (EGFR) Abrogate EGFR-Mediated Radioprotection in Non–Small Cell Lung Carcinoma

We investigate the problem of generating utter-001 ances from pairs of images showing a before 002 and an after state of a change in a visual scene. 003 We present a transformer model with differ-004 ence attention heads that learns to attend to vi-005 sual changes in consecutive images via a differ-006 ence key. We test our approach in instruction 007 generation, change captioning and difference 008 spotting and compare these tasks in terms of 009 their linguistic phenomena and reasoning abil-010 ities. Our model outperforms the state-of-the-011 art for instruction generation on the BLOCKS 012 and difference spotting on the Spot-the-diff 013 dataset and generates accurate referential and 014 compositional spatial expressions. Finally, we 015 identify linguistic phenomena that pose chal-016 lenges for generation in changing scenes. 017

Traditional methods of collecting translation and paraphrase data are prohibitively expensive, making the construction of large, new corpora difficult. While crowdsourcing offers a cheap alternative, quality control and scalability can become problematic. We discuss a novel annotation task that uses videos as the stimulus which discourages cheating. In addition, our approach requires only monolingual speakers, thus making it easier to scale since more workers are qualified to contribute. Finally, we employ a multi-tiered payment system that helps retain good workers over the long-term, resulting in a persistent, high-quality workforce. We present the results of one of the largest linguistic data collection efforts to date using Mechanical Turk, yielding 85K English sentences and more than 1k sentences for each of a dozen more languages.

Chronic postoperative inguinal pain has become a primary outcome parameter after elective inguinal hernia repair with significant consequences affecting patient productivity, employment, and quality of life. A systematic and thorough preoperative evaluation is important to identify the etiologies and types of pain. Owing to the complex nature of chronic pain, a multimodal and multidisciplinary treatment approach is recommended. Patients with chronic pain refractory to conservative measures may be considered for surgical intervention. Triple neurectomy remains the most definitive and accepted remedial operation performed and provides effective relief in the majority of patients.

e23284 Background: MEN1, DAXX and ATRX are frequently mutated in pNET. Their associations with chromosomal aberrations, disease progression, and efficacy of targeted therapy have not been established. We explored these biomarkers in patients from RADIANT-3, a randomized phase 3 trial of everolimus (EVE) vs placebo in advanced pNET (Yao JC et al, NEJM 2011). Methods: Mutations and copy number analysis was performed on 65 archival tumor samples (15% of the trial population) using next-generation sequencing. Cox PH modeling was used to assess the association between groups defined by CIN and clinical outcome. Results: MEN1, DAXX, and ATRX were mutated in 42, 28, and 11% of pNET, respectively. DAXX and ATRX mutations were mutually exclusive; while 60% of DAXX and no ATRX mutation co-exist with MEN1. Three distinct CIN patterns were identified based on 2 mutually exclusive sets of chromosome aberrations; group 1 with loss of heterozygosity (LOH) in 1st set of chromosomes; group 2 with copy-neutral LOH in 1st set and gains in 2nd set of chromosomes; and group 3 with no recurring gains or loss. MEN1/DAXX/ATRX mutations were associated with CIN in group 1 and 2 (P < 1.8 x 10-10; Table). Group 1 and 2 had better prognosis than group 3 (median overall survival, 62.6 vs 37.9 mo; HR 1.86; 95% CI 0.96-3.6). All patients derived similar PFS benefit from EVE over placebo (group 1/2: median PFS, 12.5 vs 5.4 mo; HR 0.45; 95% CI 0.21-1; group 3: median PFS, 10.8 vs 4.6 mo; HR 0.57; 95% CI 0.22-1.46), consistent to that observed in overall study population (median PFS, 11.0 vs 4.6 mo; HR 0.35; 95% CI 0.27-0.45, P< 0.001). Conclusions: pNET showed 3 distinct CIN groups, with two groups tightly associated with commonly mutated genes. These molecular subtypes may have different disease prognosis, but all benefited similarly from EVE. Results upon statistical validation in larger cohorts may have far-reaching clinical applications. Clinical trial information: NCT00510068.CIN Group N = 65 (n/%) % Samples with MEN1, DAXX or ATRX aberrations Copy number aberrations Chromosome: 1, 2, 3, 6, 8, 10, 11, 15, 16, 21, and 22 Chromosome: 4, 5, 7, 9, 12, 13, 14, 17, 18, 19, and 20 Group 1 (30/46) 97 LOH Wild- type Group 2 (5/8) 100 Copy- neutral LOH Gain Group 3 (30/46) 23 Mostly wild-type Mostly wild- type

OBJECTIVES The pharmacodynamic effects of everolimus on gastrointestinal hormone levels have not been described in patients with pancreatic neuroendocrine tumors (pNETs). We report the effects of everolimus on gastrin and glucagon levels in patients with progressive pNET in RADIANT-1 (a single-arm phase II trial) and RADIANT-3 (a placebo-controlled, randomized, phase III trial). METHODS Serum gastrin and glucagon levels were determined by immunoassay at baseline and at predose in subsequent treatment cycles in patients with elevated baseline hormone levels. The analyses included 158 patients from RADIANT-1 and 404 patients from RADIANT-3. RESULTS In RADIANT-1, everolimus induced a rapid, sustained decrease in median gastrin and glucagon levels to approximately 60% and 70% of baseline levels, respectively. In RADIANT-3, everolimus consistently reduced median gastrin and glucagon levels by greater than 50% and approximately 40%, respectively (everolimus vs placebo, P < 0.0001), whereas with placebo, both hormones at each time point were essentially the same as their baseline levels. In patients with concomitant octreotide long-acting repeatable treatment, the moderate pharmacodynamic effect on lowering gastrin was greater than that seen with everolimus alone. CONCLUSIONS In addition to prolonging progression-free survival in patients with pNET, everolimus down-regulates excess production of 2 gastrointestinal hormones, which may help control their associated clinical syndromes.

PURPOSE:Two recent phase III trials, BOLERO-1 and BOLERO-3 (Breast Cancer Trials of Oral Everolimus), evaluated the addition of everolimus to trastuzumab and chemotherapy in human epidermal growth factor receptor 2-overexpressing advanced breast cancer. The current analysis aimed to identify biomarkers to predict the clinical efficacy of everolimus treatment.METHODS:Archival tumor samples from patients in BOLERO-1 and BOLERO-3 were analyzed using next-generation sequencing, immunohistochemistry, and Sanger sequencing.RESULTS:Biomarker data were available for 549 patients. PIK3CA activating mutations and PTEN loss were reported in 30% and 16% of BOLERO-1 samples and in 32% and 12% of BOLERO-3 samples, respectively. PI3K pathway was hyperactive (PIK3CA mutations and/or PTEN loss and/or AKT1 mutation) in 47% of BOLERO-1 and 41% of BOLERO-3 samples. In both studies, differential progression-free survival (PFS) benefits of everolimus were consistently observed in patient subgroups defined by their PI3K pathway status. When analyzing combined data sets of both studies, everolimus was associated with a decreased hazard of progression in patients with PIK3CA mutations (hazard ratio [HR], 0.67; 95% CI, 0.45 to 1.00), PTEN loss (HR, 0.54; 95% CI, 0.31 to 0.96), or hyperactive PI3K pathway (HR, 0.67; 95% CI, 0.48 to 0.93). Patients with wild-type PIK3CA (HR, 1.10; 95% CI, 0.83 to 1.46), normal PTEN (HR, 1.00; 95% CI, 0.80 to 1.26), or normal PI3K pathway activity (HR, 1.19; 95% CI, 0.87 to 1.62) did not derive PFS benefit from everolimus.CONCLUSION:This analysis, although exploratory, suggests that patients with human epidermal growth factor receptor 2-positive advanced breast cancer having tumors with PIK3CA mutations, PTEN loss, or hyperactive PI3K pathway could derive PFS benefit from everolimus.

Abstract The mTORC1 inhibitor RAD001 (everolimus) has been approved for multiple cancer indications, including ER+/HER2- metastatic breast cancer. The combination of RAD001 with a dual PI3K/mTOR inhibitor BEZ235 was shown to be even more efficacious than RAD001 alone in preclinical models and was evaluated in clinical trials. We identified a male breast cancer patient who experienced a prolonged stable disease with the RAD001/BEZ235 combination as 3rd line treatment for his metastatic disease and investigated molecular mechanisms to explain the extraordinary benefit and subsequent drug resistance. The 66-year old Caucasian male had stage IIIA invasive ductal carcinoma at initial surgery. He then received two sequential adjuvant therapies: a chemo-radiotherapy and letrozole. When the patient developed multiple metastases, he was treated with chemotherapy and then fulvestrant, before received a 200 mg twice daily BEZ235 and 2.5 mg weekly RAD001 combination regimen, when a left axillary nodal metastasis was developed on fulvestrant. The patient sustained a prolonged stable disease of 18 months while under the therapy before his tumor progressed again. Tumor biopsy samples (formalin fixed, paraffin embedded) were taken from the patient at diagnosis and after progression and analyzed by immunohistochemistry (IHC) and whole exome sequencing. Blood samples were collected for germline DNA pharmacokinetic (PK) analysis. PK profile of BEZ235 was collected with Cmax, AUC, and T1/2 of 766 ng/mL, 6308 ng.h/mL, and 5.53 h, respectively. The observed BEZ235 Cmin values ranged from 75.5 to 504 ng/mL over the study. The 2.5 mg weekly RAD001 dose is substantially lower than the standard 10 mg daily dose and the Cmin could not be reliably determined. IHC assays showed low ER/PR expression (ER 30% 1+; PR 20% 1+, 10% 3+) in the diagnosis sample, but an increase of their expression (60% 1+) in the post progression sample. HER2 expression was negative in both samples. Analysis the same tumor samples using more quantitative AQUA platform demonstrated a drastic 10-fold score increase of ER from 31 to 312 and a 5-fold increase of PR from 271 to 1305. All other examined signaling pathway biomarkers (e.g. pS6, pAKT, pMAPK, pMEK and pEGFR) were expressed in both tumor samples but showed minimal expression changes between the time points. Whole exome sequencing of the diagnosis and post-progression specimens provided average coverage at 142X and 179X, respectively . No functional somatic alterations resulting in hyperactive PI3K/mTOR pathway was detected in genes such as PIK3CA, PTEN, MTOR, TSC1/2 in either tumor samples. In both specimens, a somatic deletion of 16 bp was detected in SPEN, which encodes a potential negative regulator of the estrogen signaling pathway1 . In conclusion, no PI3K/mTOR pathway hyperactivity marker was identified in the diagnostic samples. However, the increased ER/PR expression in the post-progression sample suggests that the effect of PI3K/mTOR pathway blockade by RAD001/BEZ235 might have diminished when the tumor gained further dependence on the hormonal receptor signaling. This hypothesis was supported by the observation that the patient's tumor achieved another prolonged disease control of about 13 months by exemestane, after progression from BEZ235/RAD001 treatment. 1Légaré S, et al. SPEN is a novel candidate tumor suppressor gene that regulates response to tamoxifen in estrogen receptor positive breast cancers. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013. Citation Format: A. Rose Brannron, Davide Melisi, Jennifer Hummel, Carmine Carbone, Melissa Frizziero, Wing Cheung, Parul Patel, Jorge Gallo, Giampaolo Tortora, Michael Morrissey, David Chen. Biomarker analysis of a male breast cancer patient with prolonged stable disease under mTOR/PI3K inhibitors BEZ235/RAD001. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Integrating Clinical Genomics and Cancer Therapy; Jun 13-16, 2015; Salt Lake City, UT. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(1_Suppl):Abstract nr 26.

Purpose Everolimus improved median progression-free survival by 6.4 months in patients with advanced pancreatic neuroendocrine tumors (NET) compared with placebo in the RADIANT-3 study. Here, we present the final overall survival (OS) data and data on the impact of biomarkers on OS from the RADIANT-3 study. Methods Patients with advanced, progressive, low- or intermediate-grade pancreatic NET were randomly assigned to everolimus 10 mg/day (n = 207) or placebo (n = 203). Crossover from placebo to open-label everolimus was allowed on disease progression. Ongoing patients were unblinded after final progression-free survival analysis and could transition to open-label everolimus at the investigator’s discretion (extension phase). OS analysis was performed using a stratified log-rank test in the intent-to-treat population. The baseline levels of chromogranin A, neuron-specific enolase, and multiple soluble angiogenic biomarkers were determined and their impact on OS was explored. Results Of 410 patients who were enrolled between July 2007 and March 2014, 225 received open-label everolimus, including 172 patients (85%) randomly assigned initially to the placebo arm. Median OS was 44.0 months (95% CI, 35.6 to 51.8 months) for those randomly assigned to everolimus and 37.7 months (95% CI, 29.1 to 45.8 months) for those randomly assigned to placebo (hazard ratio, 0.94; 95% CI, 0.73 to 1.20; P = .30). Elevated baseline chromogranin A, neuron-specific enolase, placental growth factor, and soluble vascular endothelial growth factor receptor 1 levels were poor prognostic factors for OS. The most common adverse events included stomatitis, rash, and diarrhea. Conclusion Everolimus was associated with a median OS of 44 months in patients with advanced, progressive pancreatic NET, the longest OS reported in a phase III study for this population. Everolimus was associated with a survival benefit of 6.3 months, although this finding was not statistically significant. Crossover of patients likely confounded the OS results.

Abstract Introduction: The prognostic and predictive utility of pretreatment serum activin A (TGF-B superfamily ligand) was correlated with the response of first-line metastatic breast cancer patients (MBC) in the phase 3 randomized trial of letrozole vs. tamoxifen. Methods: 555 ER+ first-line MBC patients had pretreatment serum available for activin A ELISA (R&D Systems). Clinical outcomes were analyzed using Cox proportional hazards modeling. Actin A levels were analyzed using continuous and categorical (median cutpoint) pretreatment serum activin A levels. A pretreatment serum activin A analysis was performed within the normal (not elevated, <15 ng/ml) pretreatment serum HER2 patient subgroup since tumor HER2 status was not available in this older clinical trial. Results: Serum activin A had a median of 971 pg/ml and an interquartile range of 623 and 1751 pg/ml. In the total population with available serum (n=555), patients with higher serum activin A (> median) had significantly reduced objective response rate (ORR)(17.33% vs 34.6%, Odds Ratio (OR)=0.4; p<0.0001) and reduced clinical benefit rate (CBR)(33% vs 53%, OR= 0.45; p<0.0001), as well as significantly shorter time to progression (TTP) [median 5.88 vs 10.98 mo, HR=1.69; p<0.0001], and reduced overall survival (OS) (median 22.78 vs 48.59 mo, HR=2.43; p<0.0001) compared to those with lower serum activin A ( In the total population with available serum (n=555), multivariate analysis for TTP revealed that high serum activin A was a significant adverse prognostic factor (HR=1.46, p<0.001). Multivariate analysis for OS also revealed that high serum activin A was an independent adverse prognostic factor (HR 1.78, p<0.0001). Outcome (Serum activin A by median cutpoint, high vs. low)TTPOSCohortPatientsHRpHRpTotal Population5551.69<0.00012.43<0.0001Letrozole Arm2741.580.0012.24<0.0001Tamoxifen Arm2611.92<0.00012.6<0.0001Serum HER2 (not elevated)3951.55<0.00012.45<0.0001 Conclusions: Patients with high pretreatment serum activin A levels had a significantly reduced ORR, CBR, TTP and OS compared to patients with low serum activin A. The results were similar within the letrozole or tamoxifen treatment arms, and within the serum HER2 not-elevated subgroups. High pretreatment serum activin A level is associated with relative resistance to hormone therapy in first-line metastatic breast cancer. Citation Format: Meghan Jensen, Ashley Kang, Suhail M Ali, Kim Leitzel, Ashwani Garg, Jaqueline Rogerio, David Chen, Raymond Hall, Scott Hofsess, Hilary A Chaudri-Ross, Nicholas Bade, Walter P Carney, Allan Lipton. Serum activin A and response to the aromatase inhibibitor (AI) letrozole versus tamoxifen in metastatic breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P3-06-14.

The present study investigated the effect of targeted mutations in the DNA-dependent protein kinase catalytic subunit and phosphorylation domains on the survival of cells in response to different qualities of ionizing radiation. Mutated Chinese hamster ovary V3 cells were exposed to 500 MeV/nucleon initial energy and 200 keV/μm monoenergetic Fe ions; 290 MeV/nucleon initial energy and average 50 keV/μm spread-out Bragg peak C ions; 70 MeV/nucleon initial energy and 1 keV/μm monoenergetic protons; and 0.663 MeV initial energy and 0.3 keV/μm Cs137 γ radiation. The results demonstrated that sensitivity to high linear energy transfer radiation is increased when both S2056 and T2609 clusters each contain a point mutation or multiple mutations are present in either cluster, whereas the phosphoinositide 3 kinase cluster only requires a single mutation to induce the sensitized phenotype of V3 cells. Additionally, the present study demonstrated that sensitivity to DNA cross-linking damage by cisplatin only requires a single mutation in one of the three clusters and that additional point mutations do not increase cell sensitivity.

PURPOSE:To explore the genetic landscape of tumors from patients enrolled on the BOLERO-2 trial to identify potential correlations between genetic alterations and efficacy of everolimus treatment. The BOLERO-2 trial has previously demonstrated that the addition of everolimus to exemestane prolonged progression-free survival by more than twofold in patients with hormone receptor-positive, human epidermal growth factor receptor 2-negative, advanced breast cancer previously treated with nonsteroidal aromatase inhibitors.PATIENTS AND METHODS:Next-generation sequencing was used to analyze genetic status of cancer-related genes in 302 archival tumor specimens from patients representative of the BOLERO-2 study population. Correlations between the most common somatic alterations and degree of chromosomal instability, and treatment effect of everolimus were investigated.RESULTS:Progression-free survival benefit with everolimus was maintained regardless of alteration status of PIK3CA, FGFR1, and CCND1 or the pathways of which they are components. However, quantitative differences in everolimus benefit were observed between patient subgroups defined by the exon-specific mutations in PIK3CA (exon 20 v 9) or by different degrees of chromosomal instability in the tumor tissues.CONCLUSION:The data from this exploratory analysis suggest that the efficacy of everolimus was largely independent of the most commonly altered genes or pathways in hormone receptor-positive, human epidermal growth factor receptor 2-negative breast cancer. The potential impact of chromosomal instabilities and low-frequency genetic alterations on everolimus efficacy warrants further investigation.

Purpose: Particle beams are increasingly applied to various cancer treatments because of their excellent dose localization to tumors while preserving surrounding normal tissues. However, characteristic of DNA damages induced by particle beams and their repair mechanisms are not fully understood. It is known that the majority of DNA double-strand breaks (DSBs) induced by ionizing radiation are repa...

The ATR (ATM [ataxia telangiectasia-mutated]- and Rad3-related) checkpoint is a crucial DNA damage signaling pathway. While the ATR pathway is known to transmit DNA damage signals through the ATR-Chk1 kinase cascade, whether post-translational modifications other than phosphorylation are important for this pathway remains largely unknown. Here, we show that protein SUMOylation plays a key role in the ATR pathway. ATRIP, the regulatory partner of ATR, is modified by SUMO2/3 at K234 and K289. An ATRIP mutant lacking the SUMOylation sites fails to localize to DNA damage and support ATR activation efficiently. Surprisingly, the ATRIP SUMOylation mutant is compromised in the interaction with a protein group, rather than a single protein, in the ATR pathway. Multiple ATRIP-interacting proteins, including ATR, RPA70, TopBP1, and the MRE11-RAD50-NBS1 complex, exhibit reduced binding to the ATRIP SUMOylation mutant in cells and display affinity for SUMO2 chains in vitro, suggesting that they bind not only ATRIP but also SUMO. Fusion of a SUMO2 chain to the ATRIP SUMOylation mutant enhances its interaction with the protein group and partially suppresses its localization and functional defects, revealing that ATRIP SUMOylation promotes ATR activation by providing a unique type of protein glue that boosts multiple protein interactions along the ATR pathway.

Abstract The BOLERO-2 phase III trial compared the combination of everolimus and exemestane to placebo and exemestane in 724 postmenopausal women with hormone receptor-positive, HER2-negative advanced breast cancer. Results showed significant improvement in progression-free survival, response rate, and clinical benefit rate. Although significant benefit has been observed in all prospectively defined subgroups, variations were seen among patients. As the first step to delineate the molecular determinants of sensitivity to everolimus and interactions between estrogen receptor and mTOR pathways, we used next-generation sequencing technology to comprehensively assess the genetic alterations in archival tumor specimens. Formalin-fixed, paraffin-embedded archival tumor samples were obtained from 496 patients. DNA was extracted from 348 samples of sufficient quantity and 230 samples (representing approximately 1/3 of all patients) qualified for the analysis. No indication of sampling bias was observed when the data were assessed and compared against several covariates as well as to the full trial data. The coding regions of 182 cancer-related genes were analyzed using an Illumina HiSeq 2000. Mutations and copy number variations were evaluated in 230 samples for which DNA extraction, library construction, and hybrid capture were successful and sequencing depth was sufficient (250-1500×). Alterations predicted to be germline events were discarded. One hundred seventy-three genes were found to be altered in at least one tumor sample. The number of alterations per sample varied from 1 to 24 (average 9 ± 4). Sequence variations (N = 1565) consisted primarily of point mutations (85%), followed by deletions (9%) and insertions (6%). 225 of 230 (98%) patients had at least two sequence variations. The most frequently mutated genes included PIK3CA (49%), TP53 (24%), and ARID1A (16%). ESR1 and IGF1R mutations were detected in 9% and 4% of the cohort, respectively. PIK3CA and AKT1 (6%) mutations were found to be mutually exclusive. Copy-number alterations comprised of 524 amplifications and 27 bi-allelic deletions. The most frequently amplified genes included CCND1 (32%) and FGFR1 (18%). Specific rearrangements assayed in 14 genes occurred infrequently (9% of the samples). These results demonstrated the feasibility of performing large-scale next-generation sequencing in a global phase III clinical trial. High alteration frequencies were observed in specific genes, resulting in activated PI3K, FGFR1, and estrogen receptor pathways, corroborating the rationale of concomitant targeting of the mTOR and estrogen receptor pathways. The results generate testable hypotheses for the development of combinations of new targeted therapies in this patient population. Citation Format: Jose Baselga, Martine Piccart, Hope Rugo, David Chen, Howard A. Burris, Mario Campone, Shinzaburo Noguchi, Alejandra Perez, Inas Deleu, Mikhail Shtivelband, Louise Provencher, Adnan Derti, Alan Huang, Rob McDonald, Creton Kalfoglou, Douglas Robinson, Tetiana Taran, Tarek Sahmoud, David Lebwohl, Gabriel N. Hortobagyi. Assessment of genetic alterations using next-generation sequencing in postmenopausal women with hormone receptor-positive, HER2-negative advanced breast cancer: results from the BOLERO-2 phase III trial. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4564. doi:10.1158/1538-7445.AM2013-4564

We previously demonstrated non-random distributions of breaks involved in chromosome aberrations induced by low- and high-LET radiation. To investigate the factors contributing to the break point distribution in radiation-induced chromosome aberrations, human epithelial cells were fixed in G1 phase. Interphase chromosomes were hybridized with a multicolor banding in situ hybridization (mBAND) probe for chromosome 3 which distinguishes six regions of the chromosome in separate colors. After the images were captured with a laser scanning confocal microscope, the 3-dimensional structure of interphase chromosome 3 was reconstructed at multimega base pair scale. Specific locations of the chromosome, in interphase, were also analyzed with bacterial artificial chromosome (BAC) probes. Both mBAND and BAC studies revealed non-random folding of chromatin in interphase, and suggested association of interphase chromatin folding to the radiation-induced chromosome aberration hotspots. We further investigated the distribution of genes, as well as the distribution of breaks found in tumor cells. Comparisons of these distributions to the radiation hotspots showed that some of the radiation hotspots coincide with the frequent breaks found in solid tumors and with the fragile sites for other environmental toxins. Our results suggest that multiple factors, including the chromatin structure and the gene distribution, can contribute to radiation-induced chromosome aberrations.

BACKGROUND:Angiomyolipomas are slow-growing tumours associated with constitutive activation of mammalian target of rapamycin (mTOR), and are common in patients with tuberous sclerosis complex and sporadic lymphangioleiomyomatosis. The insidious growth of these tumours predisposes patients to serious complications including retroperitoneal haemorrhage and impaired renal function. Everolimus, a rapamycin derivative, inhibits the mTOR pathway by acting on the mTOR complex 1. We compared the angiomyolipoma response rate on everolimus with placebo in patients with tuberous sclerosis or sporadic lymphanioleiomyomatosis-associated angiomyolipomata. METHODS:In this double-blind, placebo-controlled, phase 3 trial, patients aged 18 years or older with at least one angiomyolipoma 3 cm or larger in its longest diameter (defined by radiological assessment) and a definite diagnosis of tuberous sclerosis or sporadic lymphangioleiomyomatosis were randomly assigned, in a 2:1 fashion with the use of an interactive web response system, to receive oral everolimus 10 mg per day or placebo. The primary efficacy endpoint was the proportion of patients with confirmed angiomyolipoma response of at least a 50% reduction in total volume of target angiomyolipomas relative to baseline. This study is registered with ClinicalTrials.gov number NCT00790400. RESULTS:118 patients (median age 31·0 years; IQR 18·0–61·0) from 24 centres in 11 countries were randomly assigned to receive everolimus (n=79) or placebo (n=39). At the data cutoff, double-blind treatment was ongoing for 98 patients; two main reasons for discontination were disease progression (nine placebo patients) followed by adverse events (two everolimus patients; four placebo patients). The angiomyolipoma response rate was 42% (33 of 79 [95% CI 31–53%]) for everolimus and 0% (0 of 39 [0–9%]) for placebo (response rate difference 42% [24–58%]; one-sided Cochran-Mantel-Haenszel test p<0·0001). The most common adverse events in the everolimus and placebo groups were stomatitis (48% [38 of 79], 8% [3 of 39], respectively), nasopharyngitis (24% [19 of 79] and 31% [12 of 39]), and acne-like skin lesions (22% [17 of 79] and 5% [2 of 39]). INTERPRETATION:Everolimus reduced angiomyolipoma volume with an acceptable safety profile, suggesting it could be a potential treatment for angiomyolipomas associated with tuberous sclerosis. FUNDING:Novartis Pharmaceuticals.