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Radiation toxicities may be underestimated after treatment of transitional cell carcinoma in dogs' lower urinary tract. Assessing acute and late toxicities and differentiating them from progressive disease (PD) impacts further therapeutic approach. We retrospectively assessed dogs treated with definitive-intent chemoradiotherapy (12 × 3.8 Gy, various first-line chemotherapeutics). Local tumour control, radiation toxicities and survival were evaluated. We classified radiation toxicities according to the previously published radiation toxicity scheme "VRTOG" as well as the updated version, "VRTOG_v2.0". Fourteen dogs with transitional cell carcinoma of bladder ± urethra (n = 8), +prostate (n = 3) or solely urethra (n = 3), were included. Median follow-up was 298 days (range 185-1798 days), median overall survival 305 days (95%CI = 209;402) and 28.6% deaths were tumour-progression-related. Acute radiation toxicity was mild and self-limiting with both classification systems: In VRTOG, 5 dogs showed grade 1, and 1 dog grade 2 toxicity. In VRTOG_v2.0, 2 dogs showed grade 1, 3 dogs grade 2, and 3 dogs grade 3 toxicity. Late toxicity was noted in 14.2% of dogs (2/14) with the VRTOG, both with grade 3 toxicity. With VRTOG_v2.0, a larger proportion of 42.9% of dogs (6/14) showed late toxicities: Four dogs grade 3 (persistent incontinence), 2 dogs grade 5 (urethral obstructions without PD resulting in euthanasia). At time of death, 5 dogs underwent further workup and only 3 were confirmed to have PD. With the updated VRTOG_v2.0 classification system, more dogs with probable late toxicity are registered, but it is ultimately difficult to distinguish these from disease progression as restaging remains to be the most robust determinant.
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Carcinoma de Células Transicionales , Quimioradioterapia , Enfermedades de los Perros , Animales , Perros , Enfermedades de los Perros/terapia , Masculino , Estudios Retrospectivos , Femenino , Carcinoma de Células Transicionales/veterinaria , Carcinoma de Células Transicionales/terapia , Carcinoma de Células Transicionales/radioterapia , Carcinoma de Células Transicionales/patología , Quimioradioterapia/veterinaria , Quimioradioterapia/métodos , Quimioradioterapia/efectos adversos , Neoplasias Urológicas/veterinaria , Neoplasias Urológicas/terapia , Neoplasias Urológicas/radioterapia , Neoplasias Urológicas/patologíaRESUMEN
BACKGROUND: Classical radiation protocols are guided by physical dose delivered homogeneously over the target. Protocols are chosen to keep normal tissue complication probability (NTCP) at an acceptable level. Organs at risk (OAR) adjacent to the target volume could lead to underdosage of the tumor and a decrease of tumor control probability (TCP). The intent of our study was to explore a biology-based dose escalation: by keeping NTCP for OAR constant, radiation dose was to be maximized, allowing to result in heterogeneous dose distributions. METHODS: We used computed tomography datasets of 25 dogs with brain tumors, previously treated with 10x4 Gy (40 Gy to PTV D50). We generated 3 plans for each patient: A) original treatment plan with homogeneous dose distribution, B) heterogeneous dose distribution with strict adherence to the same NTCPs as in A), and C) heterogeneous dose distribution with adherence to NTCP <5%. For plan comparison, TCPs and TCP equivalent doses (homogenous target dose which results in the same TCP) were calculated. To enable the use of the generalized equivalent uniform dose (gEUD) metric of the tumor target in plan optimization, the calculated TCP values were used to obtain the volume effect parameter a. RESULTS: As intended, NTCPs for all OARs did not differ from plan A) to B). In plan C), however, NTCPs were significantly higher for brain (mean 2.5% (SD±1.9, 95%CI: 1.7,3.3), p<0.001), optic chiasm (mean 2.0% (SD±2.2, 95%CI: 1.0,2.8), p=0.010) compared to plan A), but no significant increase was found for the brainstem. For 24 of 25 of the evaluated patients, the heterogenous plans B) and C) led to an increase in target dose and projected increase in TCP compared to the homogenous plan A). Furthermore, the distribution of the projected individual TCP values as a function of the dose was found to be in good agreement with the population TCP model. CONCLUSION: Our study is a first step towards risk-adaptive radiation dose optimization. This strategy utilizes a biologic objective function based on TCP and NTCP instead of an objective function based on physical dose constraints.
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Neoplasias , Radioterapia de Intensidad Modulada , Humanos , Perros , Animales , Dosificación Radioterapéutica , Planificación de la Radioterapia Asistida por Computador/métodos , Radioterapia de Intensidad Modulada/métodos , Encéfalo , Probabilidad , BiologíaRESUMEN
BACKGROUND: Combined chemoradiation offers a promising therapeutic strategy for dogs with glioma. The alkylating agents temozolomide (TMZ) and lomustine (CCNU) penetrate the blood-brain barrier, and doses for dogs are established. Whether such combinations are clinically advantageous remains to be explored together with tumour-specific markers. OBJECTIVE: To investigate if triple combination of lomustine, temozolomide and irradiation reduces canine glioma cell survival in vitro. METHODS: We evaluated the sensitising effect of CCNU alone and in combination with TMZ-irradiation in canine glioma J3T-BG cells and long-term drug-exposed subclones by using clonogenic survival and proliferation assays. Bisulphite-SEQ and Western Blot were used to investigate molecular alterations. RESULTS: TMZ (200 µM) or CCNU alone (5 µM) reduced the irradiated survival fraction (4 Gy) from 60% to 38% (p = 0.0074) and 26% (p = 0.0002), respectively. The double-drug combination reduced the irradiated survival fraction (4 Gy) more potently to 12% (p < 0.0001). After long-term drug exposure, both subclones show higher IC50 values against CCNU and TMZ. For CCNU-resistant cells, both, single-drug CCNU (p = 0.0006) and TMZ (p = 0.0326) treatment combined with irradiation (4 Gy) remained effective. The double-drug-irradiation combination reduced the cell survival by 86% (p < 0.0001), compared to 92% in the parental (nonresistant) cell line. For TMZ-resistant cells, only the double-drug combination with irradiation (4 Gy) reduced the cell survival by 88% (p = 0.0057) while single-drug treatment lost efficacy. Chemoresistant cell lines demonstrated higher P-gp expression while MGMT-methylation profile analysis showed a general high methylation level in the parental and long-term treated cell lines. CONCLUSIONS: Our findings indicate that combining CCNU with TMZ-irradiation significantly reduces canine glioma cell survival. Such a combination could overcome current challenges of therapeutic resistance to improve overall patient survival.
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Enfermedades de los Perros , Glioma , Animales , Perros , Temozolomida/farmacología , Temozolomida/uso terapéutico , Lomustina/uso terapéutico , Lomustina/farmacología , Dacarbazina/farmacología , Dacarbazina/uso terapéutico , Supervivencia Celular , Glioma/veterinaria , Glioma/tratamiento farmacológico , Enfermedades de los Perros/tratamiento farmacológicoRESUMEN
Introduction: A variety of treatment options have been described for canine meningoencephalitis of unknown origin (MUO). Few studies focused on radiation therapy as a second line immunomodulating treatment, implicating its effective use. However, a standard radiation therapy protocol is lacking, and further research will help to evaluate the effect of different dose regimens. Methods: Ten dogs diagnosed with MUO based on MRI and CSF findings were prospectively enrolled. The dogs were treated with a shortened whole brain radiation therapy protocol (5 × 4 Gy) in combination with prednisolone. Neurologic changes were quantified using an established scoring scheme. Follow-up MRI and CSF examination was scheduled three months after radiation therapy. Overall survival and time to progression were calculated. Histopathology of the brain was performed in case of death. Results: Seven dogs were diagnosed de novo and three had a history of relapsing MUO. Neurological status improved in all 10 dogs during radiation therapy, with 4/10 returning to normal shortly after radiation therapy. Three dogs died within the first three months after radiation therapy. At follow-up MRI lesions completely resolved in two dogs, partially resolved in five dogs, and progressed in one dog. After follow-up MRI, dogs were further treated with prednisolone monotherapy (two dogs) and additional immunosuppressant drugs (five dogs). Overall, four dogs showed disease progression, with a mean time to progression of 691 days (95%CI: 396-987) and mean overall survival for all dogs was 723 days (95%CI: 436-1011) (both medians not reached). Histopathology confirmed MUO in three dogs but was suggestive for oligodendroglioma in one dog. Radiation induced side effects were not seen. Conclusion: Shortened whole-brain radiation therapy could be an additional treatment option for MUO in conjunction to prednisolone, specifically for cases that require rapid relief of symptoms and with relapsing history.
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OBJECTIVE: To evaluate effects of repeated alfaxalone or propofol administration on haematological and serum biochemical variables in cats undergoing radiotherapy. STUDY DESIGN: Prospective, block-randomized, clinical trial. ANIMALS: A group of 39 client-owned cats. METHODS: After butorphanol (0.2 mg kg-1) and midazolam (0.1 mg kg-1) sedation, cats were randomly assigned to receive either alfaxalone or propofol for induction of anaesthesia and sevoflurane maintenance. Cats were anaesthetized daily with the same induction agent for 10-12 days. Complete blood counts, reticulocytes, Heinz body score and serum biochemistry were performed before the first treatment (T1), at T6, T10 and 3 weeks after the final treatment (T21). Cumulative induction agent dose for each cat at each time point was evaluated for an effect on Heinz body score. Data are shown as mean ± standard deviation; p < 0.05. RESULTS: At baseline there were no significant differences in signalment or blood variables between groups. A significant decrease in haematocrit of 2.3% ± 0.77 (p = 0.02) between T1-T6 and T1-T10 [mean 4.1% (± 0.78, p < 0.0001)] was detected, with a significant increase in haematocrit of 2.1% ± 0.80 (p = 0.046) between T6-T21 and 4.0% ± 0.8 (p < 0.001) between T10-T21. Heinz body score significantly increased by 1.86 ± 0.616 (p = 0.013) between T1-T10. In the propofol group, reticulocytes increased significantly between T1-T6 [mean 23,090 µL-1 ± 7670 (p = 0.02)] and T1-T10 [mean 27,440 µL-1 ± 7990 (p = 0.007)]. Mean cumulative dose at T10 was 19.65 mg kg-1 ± 5.3 and 43.4 mg kg-1 ± 14.4 for alfaxalone and propofol, respectively, with no significant effect on Heinz body formation at any time point. CONCLUSIONS AND CLINICAL RELEVANCE: Haematocrit decreased in both groups with recovery after 3 weeks. Repeated alfaxalone and propofol administration was not associated with marked haematological or serum biochemistry changes.
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Pregnanodionas , Propofol , Gatos , Animales , Propofol/farmacología , Sevoflurano , Estudios Prospectivos , Anestesia Intravenosa/veterinaria , Pregnanodionas/farmacologíaRESUMEN
Circulating Hsp70 levels were determined in feline and porcine cohorts using two different ELISA systems. These comparative animal models of larger organisms often reflect diseases, and especially malignant tumors, better than conventional rodent models. It is therefore essential to investigate the biology and utility of tumor biomarkers in animals such as cats and pigs. In this study, levels of free Hsp70 in the blood of cats with spontaneously occurring tumors were detected using a commercial Hsp70 ELISA (R&D Systems). Sub-analysis of different tumor groups revealed that animals with tumors of epithelial origin presented with significantly elevated circulating Hsp70 concentrations. In addition to free Hsp70 levels measured with the R&D Systems Hsp70 ELISA, levels of exosomal Hsp70 were determined using the compHsp70 ELISA in pigs. Both ELISA systems detected significantly elevated Hsp70 levels (R&D Systems: median 24.9 ng/mL; compHsp70: median 44.2 ng/mL) in the blood of a cohort of APC1311/+ pigs diagnosed with high-grade adenoma polyps, and the R&D Systems Hsp70 ELISA detected also elevated Hsp70 levels in animals with low-grade polyps. In contrast, in flTP53R167H pigs, suffering from malignant osteosarcoma, the compHsp70 ELISA (median 674.32 ng/mL), but not the R&D Systems Hsp70 ELISA (median 4.78 ng/mL), determined significantly elevated Hsp70 concentrations, indicating that in tumor-bearing animals, the dominant form of Hsp70 is of exosomal origin. Our data suggest that both ELISA systems are suitable for detecting free circulating Hsp70 levels in pigs with high-grade adenoma, but only the compHsp70 ELISA can measure elevated, tumor-derived exosomal Hsp70 levels in tumor-bearing animals.
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Neoplasias Óseas , Osteosarcoma , Gatos , Animales , Porcinos , Proteínas HSP70 de Choque Térmico , Biomarcadores de Tumor , Ensayo de Inmunoadsorción Enzimática , MamíferosRESUMEN
BACKGROUND: Local progression of intracranial tumors can be the consequence of insufficient radiation dose delivered. Dose increases in the brain must be made carefully so as not to risk debilitating adverse effects such as radiation necrosis. HYPOTHESIS: A new protocol with 10 × 4 Gy + 11% physical dose increase limited to the macroscopic tumor volume results in a clinically better outcome compared to a 10 × 4 Gy protocol. ANIMALS: Fifty-seven client-owned dogs with primary intracranial neoplasia. METHODS: Randomized controlled trial. Twenty-eight dogs were assigned to the control protocol (10 × 4 Gy) and 29 to the simultaneous integrated boost (SIB) protocol with 4.45 Gy dose increase. Treatment groups were compared for outcome and signs of toxicity. RESULTS: Mild, transient acute or early-delayed adverse radiation effects were observed in 5 dogs. Severe late adverse effects were not seen. Between the protocols, no significant differences were found for outcome (intention-to-treat analysis): overall time to progression (TTP) was 708 days (95% confidence interval (95% CI) [545,872]), in the control group it was 828 days (95% CI [401,1256]), and in the SIB group 627 days (95% CI [282,973]; P = .07). Median overall survival (OS) was 684 days (95% CI [516,853]), in the control group it was 724 days (95% CI [623,826]), and in the SIB group 557 days (95% CI [95,1020]; P = .47). None of the tested variables was prognostic in terms of outcome. CONCLUSION AND CLINICAL IMPORTANCE: The dose escalation used with an 11% physical dose increase did not result in better outcome.
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Neoplasias Encefálicas , Enfermedades de los Perros , Animales , Neoplasias Encefálicas/radioterapia , Neoplasias Encefálicas/veterinaria , Ensayos Clínicos Veterinarios como Asunto , Enfermedades de los Perros/radioterapia , Perros , PronósticoRESUMEN
Tumor volume is controversially discussed as a prognostic factor in dogs treated with radiation therapy for sinonasal tumors. Dogs' body sizes vary widely and relative rather than absolute tumor volume might provide better prognostic information. Our hypothesis was that relative rather than absolute tumor volume (gross tumor volume, GTV) influences time to progression (TTP) and that a larger tumor volume is correlated with a higher tumor stage. We retrospectively investigated possible correlations of initial GTV to weight, body surface area (BSA), nasal cavity size and the tumor stage in 49 dogs with sinonasal tumors. Here, also presumed sinonasal tumors, esthesioneuroblastomas and histologically benign tumors were included. The possible impact of absolute and relative GTV on response and outcome were assessed according to imaging findings in 34 dogs with available follow-up computed tomographies (CTs) after definitive-intent radiation therapy with either a regular (10x4.2 Gy) or a simultaneously- integrated boost protocol (SIB; GTV boosted to 10x4.83 Gy). In contrast to absolute GTV (p<0.001), the relative GTVs were not correlated with dogs' body sizes. Absolute GTV, GTV relative to weight and BSA were not associated with TTP based on CT imaging. However, GTV relative to nasal cavity showed a prognostic influence with a hazard ratio of 10.97 (95%CI:1.25-96.06). When looking at GTV relative to nasal cavity, stage 3 and 4 tumors were significantly larger than stage 1 and 2 tumors (p = 0.005). Our results suggest that GTV relative to nasal cavity could be prognostic for TTP and a larger tumor volume relative to nasal cavity is correlated with a higher tumor stage.
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Neoplasias , Animales , Perros , Pronóstico , Estudios Retrospectivos , Tomografía Computarizada por Rayos X/métodos , Carga TumoralRESUMEN
PURPOSE: The FLASH effect is characterized by normal tissue sparing without compromising tumor control. Although demonstrated in various preclinical models, safe translation of FLASH-radiotherapy stands to benefit from larger vertebrate animal models. Based on prior results, we designed a randomized phase III trial to investigate the FLASH effect in cat patients with spontaneous tumors. In parallel, the sparing capacity of FLASH-radiotherapy was studied on mini pigs by using large field irradiation. EXPERIMENTAL DESIGN: Cats with T1-T2, N0 carcinomas of the nasal planum were randomly assigned to two arms of electron irradiation: arm 1 was the standard of care (SoC) and used 10 × 4.8 Gy (90% isodose); arm 2 used 1 × 30 Gy (90% isodose) FLASH. Mini pigs were irradiated using applicators of increasing size and a single surface dose of 31 Gy FLASH. RESULTS: In cats, acute side effects were mild and similar in both arms. The trial was prematurely interrupted due to maxillary bone necrosis, which occurred 9 to 15 months after radiotherapy in 3 of 7 cats treated with FLASH-radiotherapy (43%), as compared with 0 of 9 cats treated with SoC. All cats were tumor-free at 1 year in both arms, with one cat progressing later in each arm. In pigs, no acute toxicity was recorded, but severe late skin necrosis occurred in a volume-dependent manner (7-9 months), which later resolved. CONCLUSIONS: The reported outcomes point to the caveats of translating single-high-dose FLASH-radiotherapy and emphasizes the need for caution and further investigations. See related commentary by Maity and Koumenis, p. 3636.
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Carcinoma de Células Escamosas , Neoplasias Nasales , Animales , Carcinoma de Células Escamosas/patología , Gatos , Necrosis , Neoplasias Nasales/patología , Neoplasias Nasales/radioterapia , Neoplasias Nasales/veterinaria , Dosificación Radioterapéutica , Porcinos , Porcinos EnanosRESUMEN
The prognosis for canine sinonasal tumors remains rather poor despite definitive-intent radiotherapy (RT). Theoretical calculations predicted improved outcomes with simultaneously integrated boost (SIB) protocols. With the hypothesis of clinically detectable differences in outcome between groups, our retrospective study evaluated prognostic variables and outcome in dogs treated with regular versus SIB RT. Dogs with sinonasal tumors treated with either a regular (10 × 4.2 Gy) or new SIB protocol (10 × 4.83 Gy to macroscopic tumor) were included. Information regarding signalment, tumor stage, type, clinical signs, radiation toxicity, response, and outcome was collected. Forty-nine dogs were included: 27 treated regularly and 22 treated with SIB RT. A total of 69.4% showed epistaxis, 6.1% showed epileptic seizures, 46.9% showed stage IV tumors, and 6.1% showed lymph node metastases. Early toxicity was mostly mild. Late grade 1 skin toxicity (alopecia/leucotrichia) was seen in 72.1% of dogs, and a possible grade 3 ocular toxicity (blindness) was seen in one dog. Complete/partial resolution of clinical signs was seen in 95.9% of patients as best clinical response and partial remission was seen as best imaging response in 34.7%. The median progression-free survival (PFS) was 274 days (95% CI: 117-383) for regular and 300 days (95% CI: 143-451) for SIB RT, which was not significantly different (P = 0.42). Similarly, the median overall survival (OS) was 348 days (95% CI: 121-500) for regular and 381 days (95% CI: 295-634) for the SIB RT (P = 0.18). Stratified by protocol, the hazard ratio of stage IV versus stage I-III tumors was 2.29 (95% CI: 1.156-4.551, P = 0.02) for OS but not PFS. All dogs showed acceptable toxicity. In contrast to theoretical predictions, however, we could not show a statistically significant better outcome with the new protocol.
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Enfermedades de los Perros , Neoplasias , Traumatismos por Radiación , Radioterapia de Intensidad Modulada , Animales , Enfermedades de los Perros/radioterapia , Perros , Neoplasias/veterinaria , Traumatismos por Radiación/veterinaria , Dosificación Radioterapéutica/veterinaria , Radioterapia de Intensidad Modulada/veterinaria , Estudios RetrospectivosRESUMEN
A recent calculation study predicted acceptable toxicity in pelvic organs at risk for a new definitive-intent, moderately hypofractionated radiation therapy (RT) protocol (12 x 3.8 Gy), when used with image-guided intensity-modulated radiation therapy (IG-IMRT). We hypothesized this protocol to result in clinically acceptable radiation toxicities. Dogs diagnosed with and irradiated for anal sac adenocarcinoma (ASAC) were retrospectively assessed. Eleven dogs were included, six had prior surgery. Before any therapy, staging according to Polton et al. resulted in the following distribution: stage 1 (n = 1), stage 2 (n = 1), stage 3a (n = 6), stage 3b (n = 3). We scored radiation toxicities at the end of therapy, at weeks 1, 3 and every 3 months after RT according to Veterinary Radiation Therapy Oncology Group radiation toxicity criteria. Clinical follow-up was maintained on regular intervals combined with computed tomography (n = 3). Median follow-up time for dogs still alive was 594 days (range: 224-972 days). Within 1 week post treatment, eight dogs (73%) developed grade 2 and four dogs (36%) grade 1 acute toxicity in the perianal region. All acute toxicities resolved or improved to grade 1 within 3 weeks after treatment. Late toxicity, for example, chronic colitis/diarrhoea, ulcerations, strictures or myelopathies was not observed in any patient. Five dogs were euthanized 105, 196, 401, 508 and 908 days after RT and six dogs were still alive, one in spite of progressive disease. The median progression-free survival was 908 days (95%CI: 215; 1602). The previous theoretically described definitive-intent, moderately hypofractionated protocol using IG-IMRT for the treatment of advanced ASAC showed clinically acceptable acute and late toxicities.
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Adenocarcinoma , Sacos Anales , Enfermedades de los Perros , Traumatismos por Radiación , Radioterapia de Intensidad Modulada , Adenocarcinoma/radioterapia , Adenocarcinoma/veterinaria , Animales , Enfermedades de los Perros/mortalidad , Enfermedades de los Perros/radioterapia , Perros , Traumatismos por Radiación/veterinaria , Radioterapia de Intensidad Modulada/efectos adversos , Radioterapia de Intensidad Modulada/métodos , Radioterapia de Intensidad Modulada/veterinaria , Estudios RetrospectivosRESUMEN
Post-treatment outcome in canine glial tumours is described with a broad range of survival times between 2 and 28 months. After surgery or radiation therapy, the tumours may progress locally or spread within the central nervous system. It is unknown if tumour- or patient-specific factors influence prognosis. In humans, glioblastoma involving the subventricular zone has been found to recur distantly, with shortened time to progression and overall survival. We included 32 dogs irradiated for a presumptive primary glial brain tumour in this retrospective cohort study. Tumours were grouped relative to subventricular zone contact and overt ventricular invasion assessing pre-treatment magnetic resonance images. Median time to progression (TTP) for all cases was 534 days (95%CI, 310-758), with a significantly shorter TTP in dogs with lesions at the subventricular zone (median TTP, 260 vs. 687 days; p = .049). Tumours at the subventricular zone progressed more often (p = .001), and more likely as CNS-metastasis (52.9% vs. 13.3%, p = .028). Median overall survival (OS) was 489 days (95%CI, 147-831) and median tumour-specific survival 609 days (95%CI, 382-835). Involvement of the subventricular zone was significantly associated with a shorter tumour-specific survival (median, 306 vs. 719 days; p = .044). Glial tumours contacting the subventricular zone in dogs have a shorter tumour-specific survival and a higher rate of progression and CNS-metastasis. Despite local tumour control, metastasis must be considered and should prompt further treatment approaches.
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Neoplasias Encefálicas , Enfermedades de los Perros , Glioma , Animales , Neoplasias Encefálicas/patología , Neoplasias Encefálicas/radioterapia , Neoplasias Encefálicas/veterinaria , Enfermedades de los Perros/patología , Enfermedades de los Perros/radioterapia , Perros , Glioma/veterinaria , Humanos , Ventrículos Laterales/patología , Recurrencia Local de Neoplasia/patología , Recurrencia Local de Neoplasia/veterinaria , Estudios RetrospectivosRESUMEN
Intensity modulated radiation therapy (IMRT) introduced marked changes to cancer treatment in animals by reducing dose to organs at risk (OAR). As the next technological step, volumetric modulated arc therapy (VMAT) has advantages (increased degrees-of-freedom, faster delivery) compared to fixed-field IMRT. Our objective was to investigate a possible advantage of VMAT over IMRT in terms of lower OAR doses in advanced-disease sinonasal tumors in dogs treated with simultaneously-integrated boost radiotherapy. A retrospective, analytical, observational study design was applied using 10 pre-existing computed tomography datasets on dogs with stage 4 sinonasal tumors. Each dataset was planned with both, 5-field IMRT and 2 arc VMAT with 10x4.83 Gy to the gross tumor volume and 10x4.2 Gy to the planning target volume. Adequate target dose coverage and normal tissue complication probability of brain ≤5% was required. Dose constraints aspired to were D60 <15 Gy for eyes, D2 <35.4 Gy for corneae, and Dmean <20 Gy for lacrimal glands. OAR dose was statistically significantly higher in IMRT plans than in VMAT plans. Median eye D60% was 18.5 Gy (interquartile range (IQR) 17.5) versus 16.1 Gy (IQR 7.4) (p = 0.007), median lacrimal gland dose 21.8 Gy (IQR 20.5) versus 18.6 Gy (IQR 7.0) (p = 0.013), and median cornea D2% 45.5 Gy (IQR 6.8) versus 39.9 Gy (IQR 10.0) (p<0.005) for IMRT versus VMAT plans, respectively. Constraints were met in 21/40 eyes, 7/40 corneae, and 24/40 lacrimal glands. Median delivery time was significantly longer for IMRT plans than for VMAT plans (p<0.01). Based on these results, VMAT plans were found to be superior in sparing doses to eyes, lacrimal glands, corneae. However, not all ocular OAR constraints could be met while ensuring adequate dose coverage and restricting brain toxicity risk for both planning techniques.
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Neoplasias de Cabeza y Cuello , Órganos en Riesgo/efectos de la radiación , Dosificación Radioterapéutica/veterinaria , Planificación de la Radioterapia Asistida por Computador/métodos , Radioterapia de Intensidad Modulada/métodos , Animales , Perros , Neoplasias de Cabeza y Cuello/radioterapia , Neoplasias de Cabeza y Cuello/veterinaria , Proyectos Piloto , Estudios RetrospectivosRESUMEN
BACKGROUND: Similar to human glioblastoma patients, glial tumours in dogs have high treatment resistance and a guarded prognosis. In human medicine, the addition of temozolomide to radiotherapy leads to a favourable outcome in vivo as well as a higher antiproliferative effect on tumour cells in vitro. OBJECTIVES: The aim of the study was to determine the radio- and temozolomide-sensitivity of three canine glial tumour cell lines and to investigate a potential additive cytotoxic effect in combined treatment. Additionally, we wanted to detect the level of MGMT promoter methylation in these cell lines and to investigate a potential association between MGMT promoter methylation and treatment resistance. METHODS: Cells were treated with various concentrations of temozolomide and/or irradiated with 4 and 8 Gy. Radiosensitization by temozolomide was evaluated using proliferation assay and clonogenic assay, and MGMT DNA methylation was investigated using bisulfite next-generation sequencing. RESULTS: In all tested canine cell lines, clonogenicity was inhibited significantly in combined treatment compared to radiation alone. All canine glial cell lines tested in this study were found to have high methylation levels of MGMT promoter. CONCLUSIONS: Hence, an additive effect of combined treatment in MGMT negative canine glial tumour cell lines in vitro was detected. This motivates to further investigate the association between treatment resistance and MGMT, such as MGMT promoter methylation status.
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Neoplasias Encefálicas , Enfermedades de los Perros , Glioma , Animales , Neoplasias Encefálicas/tratamiento farmacológico , Neoplasias Encefálicas/radioterapia , Neoplasias Encefálicas/veterinaria , Línea Celular , Metilasas de Modificación del ADN/genética , Metilasas de Modificación del ADN/metabolismo , Enzimas Reparadoras del ADN/genética , Enzimas Reparadoras del ADN/metabolismo , Dacarbazina/farmacología , Enfermedades de los Perros/tratamiento farmacológico , Enfermedades de los Perros/radioterapia , Perros , Glioma/tratamiento farmacológico , Glioma/radioterapia , Glioma/veterinaria , Humanos , Temozolomida/farmacología , Proteínas Supresoras de Tumor/genética , Proteínas Supresoras de Tumor/metabolismoRESUMEN
BACKGROUND: Rodent cancer models have limitations in predicting efficacy, tolerability and accompanying biomarkers of ICIs in humans. Companion dogs suffering from neoplastic diseases have gained attention as a highly relevant translational disease model. Despite successful reports of PD-1/PD-L1 blockade in dogs, no compounds are available for veterinary medicine. METHODS: Here, we assessed suitability of seven FDA-approved human ICIs to target CTLA-4 or PD-1/PD-L1 in dogs. Cross-reactivity and blocking potential was assessed using ELISA and flow cytometry. Functional responses were assessed on peripheral blood mononuclear cells (PBMCs) derived from healthy donors (n = 12) and cancer patient dogs (n = 27) as cytokine production after stimulation. Immune composition and target expression of healthy donors and cancer patients was assessed via flow cytometry. RESULTS: Four candidates showed cross-reactivity and two blocked the interaction of canine PD-1 and PD-L1. Of those, only atezolizumab significantly increased cytokine production of healthy and patient derived PBMCs in vitro. Especially lymphoma patient PBMCs responded with increased cytokine production. In other types of cancer, response to atezolizumab appeared to correlate with a lower frequency of CD8 T cells. CONCLUSIONS: Cross-functionality of atezolizumab encourages reverse translational efforts using (combination) immunotherapies in companion dog tumor patients to benefit both veterinary and human medicine.
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Planning organ at risk volume (PRV) estimates have been reported as methods for sparing organs at risk (OARs) during radiation therapy, especially for hypofractioned and/or dose-escalated protocols. The objectives of this retrospective, analytical, observational study were to evaluate peri-ocular OAR shifts and derive PRVs in a sample of dogs undergoing radiation therapy for periocular tumors. Inclusion criteria were as follows: dogs irradiated for periocular tumors, with 3D-image-guidance and at least four cone-beam CTs (CBCTs) used for position verification, and positioning in a rigid bite block immobilization device. Peri-ocular OARs were contoured on each CBCT and the systematic and random error of the shifts in relation to the planning CT position computed. The formula 1.3×Σ+0.5xσ was used to generate a PRV of each OAR in the dorsoventral, mediolateral, and craniocaudal axis. A total of 30 dogs were sampled, with 450 OARs contoured, and 2145 shifts assessed. The PRV expansion was qualitatively different for each organ (1-4 mm for the dorsoventral and 1-2 mm for the mediolateral and craniocaudal axes). Maximal PRV expansion was ≤4 mm and directional for the majority; most pronounced for corneas and retinas. Findings from the current study may help improve awareness of and minimization of radiation dose in peri-ocular OARs for future canine patients. Because some OARs were difficult to visualize on CBCTs and/ or to delineate on the planning CT, authors recommend that PRV estimates be institution-specific and applied with caution.
Asunto(s)
Tomografía Computarizada de Haz Cónico/veterinaria , Imagenología Tridimensional/veterinaria , Planificación de la Radioterapia Asistida por Computador/veterinaria , Animales , Perros , Tamaño de los Órganos , Órganos en Riesgo/efectos de la radiación , Planificación de la Radioterapia Asistida por Computador/métodos , Radioterapia Guiada por Imagen/métodos , Radioterapia Guiada por Imagen/veterinaria , Estudios RetrospectivosRESUMEN
Image-guided, intensity modulated radiation therapy (IG-IMRT) reduces dose to pelvic organs at risk without losing dose coverage to the planning target volume (PTV) and might permit margin reductions potentially resulting in lower toxicity. Appropriate PTV margins have not been established for IG-IMRT in abdominopelvic tumours in dogs, and herein we explore if our usual PTV 5 mm margin can be reduced further. Datasets from dogs that underwent IG-IMRT for non-genitourinary abdominopelvic neoplasia with 5 mm-PTV expansion were included in this retrospective virtual study. The clinical target volumes and organs at risk (OAR) colon, rectum, spinal cord were adapted to each co-registered cone-beam computed tomography (CBCT) used for positioning. New treatment plans were generated and smaller PTV margins of 3 mm and 4 mm evaluated with respect to adequate dose coverage and normal tissue complication probability (NTCP) of OAR. Ten dogs with a total of 70 CBCTs were included. Doses to the OAR of each CBCT deviated mildly from the originally planned doses. In some plans, insufficient build-up of the high dose-area at the body surface was found due to inadequate or missing bolus placement. Overall, the margin reduction to 4 mm or 3 mm did not impair dose coverage and led to significantly lower NTCP in all OAR except for spinal cord delayed myelopathy. However, overall NTCP for spinal cord was very low (<4%). PTV-margins depend on patient immobilization and treatment technique and accuracy. IG-IMRT allows treatment with very small margins in the abdominopelvic region, ensuring appropriate target dose coverage, while minimizing NTCP.
Asunto(s)
Enfermedades de los Perros , Neoplasias de la Próstata , Radioterapia de Intensidad Modulada , Animales , Enfermedades de los Perros/radioterapia , Perros , Masculino , Probabilidad , Neoplasias de la Próstata/veterinaria , Dosificación Radioterapéutica/veterinaria , Planificación de la Radioterapia Asistida por Computador/veterinaria , Radioterapia de Intensidad Modulada/efectos adversos , Radioterapia de Intensidad Modulada/veterinaria , Estudios RetrospectivosRESUMEN
In this work, a method is established to calibrate a model that describes the basic dynamics of DNA damage and repair. The model can be used to extend planning for radiotherapy and hyperthermia in order to include the biological effects. In contrast to "syntactic" models (e.g., describing molecular kinetics), the model used here describes radiobiological semantics, resulting in a more powerful model but also in a far more challenging calibration. Model calibration is attempted from clonogenic assay data (doses of 0-6 Gy) and from time-resolved comet assay data obtained within 6 h after irradiation with 6 Gy. It is demonstrated that either of those two sources of information alone is insufficient for successful model calibration, and that both sources of information combined in a holistic approach are necessary to find viable model parameters. Approximate Bayesian computation (ABC) with simulated annealing is used for parameter search, revealing two aspects that are beneficial to resolving the calibration problem: (1) assessing posterior parameter distributions instead of point-estimates and (2) combining calibration runs from different assays by joining posterior distributions instead of running a single calibration run with a combined, computationally very expensive objective function.
Asunto(s)
Supervivencia Celular , Daño del ADN , Modelos Biológicos , Animales , Teorema de Bayes , Línea Celular Tumoral , Supervivencia Celular/efectos de la radiación , Ensayo Cometa , Biología Computacional , Reparación del ADN , Perros , Humanos , Conceptos Matemáticos , Método de Montecarlo , Neoplasias/metabolismo , Neoplasias/patología , Neoplasias/radioterapia , Planificación de la Radioterapia Asistida por Computador , Biología de Sistemas , Ensayo de Tumor de Célula MadreRESUMEN
Pre-treatment of tumors with hyperthermia is often used to increase the efficacy of radiotherapy. One of the main proteins induced in response to hyperthermia is heat shock protein 70 (HSP70). The aim of our study was to investigate up- and down-regulated genes in response to (thermo)radiotherapy in HSP70 proficient and deficient canine osteosarcoma cell line (Abrams), and functional role of HSP70 in the mechanism of thermoradiosensitization. Cells were transfected with negative control siRNA or siRNA targeting HSP70 and treated with hyperthermia (HT), radiotherapy (RT), and thermoradiotherapy (HTRT). RNA sequencing was used to analyze gene expression. Hyperthermia and thermoradiotherapy, but not radiotherapy alone, induced differential gene expression. We identified genes differentially expressed only in HSP70 knockdown (thus HSP70-dependent) cells in response to hyperthermia and thermoradiotherapy. Interestingly, cell proliferation but not clonogenicity and apoptosis/necrosis was affected by the HSP70 knockdown in response to thermoradiotherapy. The results suggest that HSP70 regulates expression of specific genes in response to hyperthermia and thermoradiotherapy. Further investigations into the role of specific genes regulated in a HSP70-dependent manner in response to thermoradiotherapy could pave a way into new, combinatorial treatment options for (canine) osteosarcoma and other cancer types.