Circulating tumor DNA tracking through driver mutations as a liquid biopsy-based biomarker for uveal melanoma

© The Author(s). 2021 Open Access. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.Background: Uveal melanoma (UM) is the most common intraocular tumor in adults. Despite good primary tumor control, up to 50% of patients develop metastasis, which is lethal. UM often presents asymptomatically and is usually diagnosed by clinical examination and imaging, making it one of the few cancer types diagnosed without a biopsy. Hence, alternative diagnostic tools are needed. Circulating tumor DNA (ctDNA) has shown potential as a liquid biopsy target for cancer screening and monitoring. The aim of this study was to evaluate the feasibility and clinical utility of ctDNA detection in UM using specific UM gene mutations. Methods: We used the highly sensitive digital droplet PCR (ddPCR) assay to quantify UM driver mutations (GNAQ, GNA11, PLCβ4 and CYSTLR2) in cell-free DNA (cfDNA). cfDNA was analyzed in six well established human UM cell lines with known mutational status. cfDNA was analyzed in the blood and aqueous humor of an UM rabbit model and in the blood of patients. Rabbits were inoculated with human UM cells into the suprachoroidal space, and mutated ctDNA was quantified from longitudinal peripheral blood and aqueous humor draws. Blood clinical specimens were obtained from primary UM patients (n = 14), patients presenting with choroidal nevi (n = 16) and healthy individuals (n = 15). Results: The in vitro model validated the specificity and accuracy of ddPCR to detect mutated cfDNA from UM cell supernatant. In the rabbit model, plasma and aqueous humor levels of ctDNA correlated with tumor growth. Notably, the detection of ctDNA preceded clinical detection of the intraocular tumor. In human specimens, while we did not detect any trace of ctDNA in healthy controls, we detected ctDNA in all UM patients. We observed that UM patients had significantly higher levels of ctDNA than patients with nevi, with a strong correlation between ctDNA levels and malignancy. Noteworthy, in patients with nevi, the levels of ctDNA highly correlated with the presence of clinical risk factors. Conclusions: We report, for the first time, compelling evidence from in vitro assays, and in vivo animal model and clinical specimens for the potential of mutated ctDNA as a biomarker of UM progression. These findings pave the way towards the implementation of a liquid biopsy to detect and monitor UM tumors.info:eu-repo/semantics/publishedVersio

Treatment with tocilizumab or corticosteroids for COVID-19 patients with hyperinflammatory state: a multicentre cohort study (SAM-COVID-19)

Objectives: The objective of this study was to estimate the association between tocilizumab or corticosteroids and the risk of intubation or death in patients with coronavirus disease 19 (COVID-19) with a hyperinflammatory state according to clinical and laboratory parameters. Methods: A cohort study was performed in 60 Spanish hospitals including 778 patients with COVID-19 and clinical and laboratory data indicative of a hyperinflammatory state. Treatment was mainly with tocilizumab, an intermediate-high dose of corticosteroids (IHDC), a pulse dose of corticosteroids (PDC), combination therapy, or no treatment. Primary outcome was intubation or death; follow-up was 21 days. Propensity score-adjusted estimations using Cox regression (logistic regression if needed) were calculated. Propensity scores were used as confounders, matching variables and for the inverse probability of treatment weights (IPTWs). Results: In all, 88, 117, 78 and 151 patients treated with tocilizumab, IHDC, PDC, and combination therapy, respectively, were compared with 344 untreated patients. The primary endpoint occurred in 10 (11.4%), 27 (23.1%), 12 (15.4%), 40 (25.6%) and 69 (21.1%), respectively. The IPTW-based hazard ratios (odds ratio for combination therapy) for the primary endpoint were 0.32 (95%CI 0.22-0.47; p < 0.001) for tocilizumab, 0.82 (0.71-1.30; p 0.82) for IHDC, 0.61 (0.43-0.86; p 0.006) for PDC, and 1.17 (0.86-1.58; p 0.30) for combination therapy. Other applications of the propensity score provided similar results, but were not significant for PDC. Tocilizumab was also associated with lower hazard of death alone in IPTW analysis (0.07; 0.02-0.17; p < 0.001). Conclusions: Tocilizumab might be useful in COVID-19 patients with a hyperinflammatory state and should be prioritized for randomized trials in this situatio

Disseminated uveal melanoma : the seeds of metastases

Treatment of primary uveal melanoma (UM) appears to have no direct influence on the evolution of metastatic disease, as 50% of patients will die of metastases following successful local therapy. This fact suggests that undiagnosed malignant cells have already disseminated at the time of initial treatment. The purpose of the study was to detect early stages of tumor cell dissemination in the form of circulating malignant cells (CMCs) in UM patients during a prospective longitudinal study.CMCs were detected using nested RT-PCR with two markers: tyrosinase and Melan A. These markers were found in our first transitional study to be highly expressed in 46 cases of UM, as well as in CMCs and metastatic sites of our UM animal model.The RT-PCR technique was developed and evaluated in a second transitional study. Blood from 60 healthy volunteers was spiked with increasing concentrations of UM cells and evaluated with double-marker nested RT-PCR. The technique is highly specific (100%) with 70% sensitivity for Tyrosinase and 72% for Melan A. Further improvement on sensitivity was achieved by running multiple (n=10) RT-PCR per sample. With this approach, all samples tested positive regardless of the number of cell spiked, with no compromise in final test specificity.Finally, we identified CMCs in 29 of 30 UM patients. A total of 136 samples were collected during 18 months, with 2720 RT-PCR (10 RT-PCR per sample) performed. 119 samples (87.5%) tested positive; however, a low number of positive tests were recorded per sample (2 of 10 for Melan A and 1 of 10 for tyrosinase). CMCs were found in newly diagnosed, irradiated, enucleated, and observed patients regardless of tumor size at initial diagnosis and the time interval following treatment.In conclusion, we established that CMCs are present in the majority of UM patients. CMCs were detected at time of initial primary tumor diagnosis as well as many months to years following local treatment. As the survival of CMCs in blood in known to be limited, a source of replicating cells is necessary to maintain their longitudinal presence in blood. The intraocular tumor may be a first early source of CMCs. We can hypothesize that micrometastatic foci would become a secondary source of CMCs following primary tumor treatment. CMCs may extravasate and remain dormant, in a state in which replication may be balanced with cell release into the circulation, with no evidence of progressive tumor growth. The organs involved in tumor dormancy, the triggers that stimulate dormant cells to form macrometastases, and the appropriate patient selection for adjuvant therapy remain as attractive areas for future research

Conjunctival Involvement of T-Cell Lymphoma in a Patient with Mycosis Fungoides

Background. Ocular involvement in mycosis fungoides (MF) cases occurs in one-third of patients with the eyelid being the most frequent site affected; however, conjunctival involvement is rarely reported. Herein, we report a rare case of conjunctival involvement of MF. Case Presentation. A 66-year-old man who was previously diagnosed with MF in 2010 and was treated presented in 2014 complaining of foreign body sensation and redness in both eyes. Slit lamp examination of both eyes showed erythematous conjunctival growth that extended circumferentially. Physical examination revealed erythematous skin lesions on different body parts. Conjunctival biopsy was performed and revealed a dense, highly polymorphic lymphocytic population. The immunophenotype demonstrated a neoplastic T-cell origin consistent with MF. A diagnosis of conjunctival involvement by MF was made. The conjunctiva was treated with radiotherapy resulting in tumor regression. There were no recurrences at the 6-month follow-up. Conclusion. T-cell lymphoma should be considered in patients with a history of MF presenting with conjunctival and skin lesions

Calidad de vida relacionada con la salud y estrategias de afrontamiento ante el dolor en pacientes atendidos por una unidad de tratamiento del dolor

Introduction: The World Health Organisation (WHO) defines the quality of life (QOL) as a subjective evaluation of the characteristics of a person life, a composite variable referring to an individual's subjective overall satisfaction with life, a multidimensional construct primarily based on a person's subjective appraisal of their physical, functional, emotional and social well-being. Therefore health-related quality of life (HRQOL) refers to the extent to which one's usual or expected physical, emotional and social well-being are affected by a medical condition or its treatment. Objectives: Describe the different pathology of chronic no malign pain in patients who go for the first time to Unit of pain during 6 months and the treatment receive. Know the changes in health-related quality of life (HRQL), pain intensity, coping strategies and anxiety/depression after six months. Know the relation between HRQL, coping strategies, anxiety/depression and pain intensity in these patients. Know the changes in valúes which measures previous parameters in depending of diagnostic groups. Material and methods: This work is descriptive with measures repeated after 6 months about the population of patients that go to a Unit of pain Galdakao's Hospital during the second part of 2005. Several questionnaires are administrated about quality of life (SF-36), coping strategies (CAD-R), anxiety/depression (HAD) and intensity of pain (EVA). The questionnaire SF-36, CAD-R, HAD, and EVA were administered again and were collected the different treatment given by unit of pain. All proceedings were realized with the statistics packet SAS System v9.1, we assumed statistic signification with p< 0.05. Results and conclusions: The sample is composed of 119 patients, 93 completed the questionnaires, at the beginning and 6 months after, so we have 21.8% lost. In the SF-36 the improvement is significative in domains role physical, corporal pain, vitality, mental health and the standard physical component. In the HAD anxiety values, increase and depression values decrease in a no significative mode. Pain intensitivity is modérate, decreases in a no significative mode. Coping strategies more used are autoafirmation and the look for information, 6 months after, coping strategies active are more used. We can see the negative association between quality of life with anxiety and depression and pain intensitivity. The worst group responding to treatment by Unit of Pain is fibromyalgia patients.Introducción: La Organización Mundial de la Salud, define la calidad de vida como: "la percepción del individuo de su situación en la vida, dentro del contexto cultural y de valores en el que vive, y en relación con sus objetivos, expectativas, valores e intereses". Por lo tanto para una valoración completa de los beneficios producidos por un determinado tratamiento, es esencial medir su impacto en el estado de salud del paciente, lo que se denomina calidad de vida relacionada con la salud (CVRS). Objetivos: Describir las diferentes patologías de dolor crónico no maligno de los pacientes que acuden por primera vez a una unidad del dolor (UTD) a lo largo de 6 meses y el tratamiento recibido. Conocer los cambios en la CVRS, intensidad del dolor, estrategias de afrontamiento y ansiedad/depresión al cabo de 6 meses de seguimiento de estos pacientes. Conocer la relación entre CVRS, afrontamiento, ansiedad/depresión e intensidad del dolor en estos pacientes. Conocer los cambios en las puntuaciones que miden los parámetros anteriores según los grupos diagnósticos. Material y métodos: Estudio descriptivo con medidas repetidas a los 6 meses respecto a la población de pacientes que acuden a la UTD del Hospital de Galdakao (Bizkaia) durante el 2º semestre del 2005. Se administran varios cuestionarios, de calidad de vida (SF-36), de afrontamiento (CAD-R), ansiedad y depresión (HAD) y la intensidad del dolor (EVA). Al cabo de 6 meses se vuelven a administrar y se recogen los diferentes tratamientos realizados en la UTD. Todos los procedimientos fueron realizados con el paquete estadístico SAS System v9.1, se asumió significación estadística cuando p < 0.05. Resultados y conclusiones: La muestra la conforman 119 pacientes, de ellos 93 completaron los cuestionarios al inicio del estudio y al cabo de 6 meses, por lo que presentamos un 21.85 % de pérdidas. Los resultados del estudio muestran que los pacientes refieren una mejoría significativa en los dominios rol físico, dolor corporal y vitalidad del SF-36 al cabo de 6 meses de su paso por la UTD. Se observa una tendencia a que disminuya la intensidad del dolor y la sintomatología depresiva, sin embargo la sintomatología ansiosa tiende a aumentar. Las estrategias de afrontamiento más utilizadas por la muestra son las activas. Existe una asociación negativa entre el estado de salud con la sintomatología de ansiedad, depresión y la intensidad del dolor. Dentro de los grupos diagnósticos el grupo de fibromialgia y dolor osteoarticular es el que peor evoluciona, tanto en parámetros de intensidad de dolor, como en sintomatología ansiosa y depresiva

Trends in Radiation Practices for Female Ocular Oncologists in North America: A Collaborative Study of the International Society of Ocular Oncology

The aim of this study was to determine the known radiation exposure, attitudes, and consequent risk modifications among female ocular oncologists in North America who routinely administer radioactive plaque brachytherapy treatment and are members of the International Society of Ocular Oncology. Nineteen female ocular oncologists completed an anonymous 17-question radiation exposure survey. Eleven of the participants chose to routinely wear lead protection during surgery; 8 did not. Fifteen of 19 participants reported using an unloaded "nonactive" template to prepare for plaque implantation. During pregnancy, 11 of 13 participants continued to perform plaque brachytherapy. Eight of these 11 undertook measures to decrease radiation exposure self-reported as lead wear and other. The average reported anxiety regarding fertility was 2.1 (SD, 2.2) on a scale from 1 to 10. This study corroborates prior literature that surgeons' exposure to radiation during plaque brachytherapy is minimal. Nonetheless, there remains some anxiety regarding exposure risk to women, due to potential effects on fertility and fetal health. We found variability in exposure monitoring, required training, and precautions during pregnancy amongst this group of surgeons. Improved education and clearer pregnancy guidelines may equip female ocular oncologists with optimal knowledge regarding risk of radiation exposure

Aps Sustainable and Inclusive Social Volunteering: learning by doing a service stop the community (II)

El objetivo principal es consolidar el desarrollo de competencias genéricas y específicas que capacite a los y las participantes para transmitir la importancia de la Agenda 2030 en la universidad y la comunidad, desarrollando capacidades de reafirmación social para canalizar acciones encaminadas hacia los Objetivos de Desarrollo Sostenible (ODS). A través de la metodología del Aprendizaje-Servicio se busca promover un encuentro para el trabajo conjunto, mediante la conexión entre la individualidad de las personas, el trabajo grupal y comunitario. Los fundamentos del Trabajo Social nos permitirán trabajar un enfoque de derechos, valores y compromiso. Además, se promoverá el desarrollo de habilidades para la vida desde la educación popular, la educación eco-social, el diseño universal del aprendizaje, los entornos inclusivos, la lectura fácil y los huertos eco-didácticos. En el proyecto participará un grupo de veinte voluntarios/as (estudiantes de grado y máster de la Facultad de Trabajo Social y personas externas con diversidad intelectual), puntualmente estudiantes de Bellas Artes y Estudios Estadísticos. Como Aprendizaje, las actividades formativas programadas buscan formar al voluntariado tanto en los ODS como en las capacidades comentadas anteriormente. Para su desarrollo, se contará con recursos de las entidades colaboradoras, tanto propias de la UCM como externas. Como Servicio a la Comunidad, se propondrá al voluntariado el reto de ser embajadores/as de los ODS, compartiendo su mensaje en la universidad y comunidad, desde el conocimiento científico y el reconocimiento a los demás (cultura, saberes y procesos distintos a los nuestros). La gestión operativa se realizará a tres niveles: equipo técnico, para el seguimiento y evaluación del proyecto; equipo de acompañamiento, para coordinar actividades con el voluntariado y garantizar cohesión grupal; y equipo de relaciones institucionales, para coordinar con entidades colaboradoras los espacios y agenda para las actividades con el voluntariado.The main objective is to consolidate the development of generic and specific skills that train the participants to convey the importance of the 2030 Agenda in the university and the community, developing capacities for social reaffirmation to channel actions aimed at the Sustainable Development Goals ( ODS). Through the Service-Learning methodology, it seeks to promote a meeting for joint work, through the connection between the individuality of people, group and community work. The foundations of Social Work will allow us to work on a focus on rights, values ​​and commitment. In addition, the development of life skills will be promoted from popular education, eco-social education, universal learning design, inclusive environments, easy reading and eco-didactic gardens. A group of twenty volunteers will participate in the project (undergraduate and master's students from the Faculty of Social Work and external people with intellectual diversity), occasionally students of Fine Arts and Statistical Studies. As Learning, the programmed training activities seek to train volunteers both in the SDGs and in the capacities mentioned above. For its development, there will be resources from collaborating entities, both from the UCM and from outside. As Community Service, the challenge of being ambassadors of the SDGs will be proposed to volunteers, sharing their message in the university and community, based on scientific knowledge and recognition of others (culture, knowledge and processes different from ours). Operational management will be carried out at three levels: technical team, for the monitoring and evaluation of the project; support team, to coordinate activities with volunteers and guarantee group cohesion; and institutional relations team, to coordinate with collaborating entities the spaces and agenda for volunteer activities.Depto. de Trabajo Social y Servicios SocialesFac. de Trabajo SocialTRUEServicio ApS. Universidad Complutense de Madridunpu