Engineered CAR-T cells targeting the non-functional P2X purinoceptor 7 (P2X7) receptor as a novel treatment for ovarian cancer.

Objectives: Recent studies have identified expression of the non-functional P2X7 (nfP2X7) receptor on various malignant cells including ovarian cancer, but not on normal cells, which makes it a promising tumour-associated antigen candidate for chimeric antigen receptor (CAR)-T-cell immunotherapies. In this study, we assessed the cytotoxic effects of nfP2X7-CAR-T cells on ovarian cancer using in vitro and in vivo models. Methods: We evaluated the effects of nfP2X7-CAR-T cells on ovarian cancer cell lines (SKOV-3, OVCAR3, OVCAR5), normal peritoneal cells (LP-9) and primary serous ovarian cancer cells derived from patient ascites in vitro using monolayer and 3D spheroid assays. We also evaluated the effects of nfP2X7-CAR-T cells on patient-derived tissue explants, which recapitulate an intact tumour microenvironment. In addition, we investigated the effect of nfP2X7-CAR-T cells in vivo using the OVCAR-3 xenograft model in NOD-scid IL2Rγnull (NSG) mice. Results: Our study found that nfP2X7-CAR-T cells were cytotoxic and significantly inhibited survival of OVCAR3, OVCAR5 and primary serous ovarian cancer cells compared with un-transduced CD3+ T cells in vitro. However, no significant effects of nfP2X7-CAR-T cells were observed for SKOV3 or normal peritoneal cells (LP-9) cells with low P2X7 receptor expression. Treatment with nfP2X7-CAR-T cells increased apoptosis compared with un-transduced T cells in patient-derived explants and correlated with CD3 positivity. Treatment with nfP2X7-CAR-T cells significantly reduced OVCAR3 tumour burden in mice compared with un-transduced CD3 cells for 7-8 weeks. Conclusion: This study demonstrates that nfP2X7-CAR-T cells have great potential to be developed as a novel immunotherapy for ovarian cancer.

Functional assessment of children and adolescents with autism spectrum disorder in Argentina: ICF-ASD multicenter study. / Evaluación funcional de niños, niñas y adolescentes con diagnóstico de trastorno del espectro autista en Argentina: estudio multicéntrico TEA-CIFunciona.

Introduction. Autism spectrum disorder (ASD) is characterized by difficulties in social communication and repetitive and stereotyped behaviors. In addition to the diagnostic category, the activities performed by children and adolescents and their social involvement are the main aspects to be considered according to the International Classification of Functioning, Disability, and Health (ICF) proposed by the World Health Organization to describe health status. In a previous study, we developed the first version of a pediatric tool based on the ICF called ICF-ASD for the functional assessment of children and adolescents with ASD to capture functional characteristics adapted to our cultural setting. Our subsequent objective was to apply the ICF-ASD in a multicenter format to assess children and adolescents from different regions, review, and update it, and identify barriers and facilitators. Population and methods. The ICF-ASD version 1.0 was administered to children and adolescents younger than 16 years with a confirmed diagnosis of ASD (as per DSM-5 criteria), who were receiving follow-up at 5 children's health centers across Argentina. Results. Version 2.0 of the ICF-ASD was obtained, which included 34 categories (10 under body function, 15 under activities and participation, and 9 under environmental factors). A functional profile was developed for the whole sample (n = 308). Conclusions. The updated version of the ICF-ASD helps to standardize and systematize the collection of necessary data for an adequate follow-up of children and adolescents with ASD at a national level. It also allows to identify barriers to overcome and facilitators to be generalized.

Introducción. El trastorno del espectro autista (TEA) se caracteriza por dificultades de comunicación social y comportamientos repetitivos y estereotipados. Además de la categoría diagnóstica, las actividades que los niños, niñas y adolescentes (NNyA) pueden realizar y la participación social son los aspectos principales por considerar desde el marco de la Clasificación Internacional del Funcionamiento, la Discapacidad y la Salud (CIF), propuesta por la Organización Mundial de la Salud, para describir los estados de salud. En una investigación previa, elaboramos la primera versión de una herramienta pediátrica basada en la CIF llamada TEA-CIFunciona para evaluación funcional de NNyA con diagnóstico de TEA, que permitió captar características funcionales adaptadas a nuestro contexto cultural. Se propuso como objetivo posterior aplicar TEA-CIFunciona en formato multicéntrico para evaluar NNyA de diferentes regiones, revisar y actualizar la herramienta, e identificar barreras y facilitadores. Población y métodos. Se administró TEA-CIFunciona versión 1.0 a NNyA con diagnóstico confirmado de TEA (según criterios del DSM-5), menores de 16 años, en seguimiento en cinco centros de atención pediátrica del país. Resultados. Se obtuvo la versión 2.0 de TEA-CIFunciona con 34 categorías (10 funciones corporales, 15 actividades y participación, y 9 factores ambientales). Se elaboró el perfil funcional de la muestra completa (n = 308). Conclusiones. La versión actualizada de TEA-CIFunciona contribuye a estandarizar y a sistematizar la obtención de información necesaria para adecuar el seguimiento de los NNyA con TEA a nivel nacional. Además, permite identificar barreras por superar y facilitadores para generalizar.

Pre-clinical validation of a pan-cancer CAR-T cell immunotherapy targeting nfP2X7.

Chimeric antigen receptor (CAR)-T cell immunotherapy is a novel treatment that genetically modifies the patients' own T cells to target and kill malignant cells. However, identification of tumour-specific antigens expressed on multiple solid cancer types, remains a major challenge. P2X purinoceptor 7 (P2X7) is a cell surface expressed ATP gated cation channel, and a dysfunctional version of P2X7, named nfP2X7, has been identified on cancer cells from multiple tissues, while being undetectable on healthy cells. We present a prototype -human CAR-T construct targeting nfP2X7 showing potential antigen-specific cytotoxicity against twelve solid cancer types (breast, prostate, lung, colorectal, brain and skin). In xenograft mouse models of breast and prostate cancer, CAR-T cells targeting nfP2X7 exhibit robust anti-tumour efficacy. These data indicate that nfP2X7 is a suitable immunotherapy target because of its broad expression on human tumours. CAR-T cells targeting nfP2X7 have potential as a wide-spectrum cancer immunotherapy for solid tumours in humans.

Inertial Microfluidic Purification of CAR-T-Cell Products.

Chimeric antigen receptor T (CAR-T) cell therapy is rapidly becoming a frontline cancer therapy. However, the manufacturing process is time-, labor- and cost-intensive, and it suffers from significant bottlenecks. Many CAR-T products fail to reach the viability release criteria set by regulators for commercial cell therapy products. This results in non-recoupable costs for the manufacturer and is detrimental to patients who may not receive their scheduled treatment or receive out-of-specification suboptimal formulation. It is demonstrated here that inertial microfluidics can, within minutes, efficiently deplete nonviable cells from low-viability CAR-T cell products. The percentage of viable cells increases from 40% (SD ± 0.12) to 71% (SD ± 0.09) for untransduced T cells and from 51% (SD ± 0.12) to 71% (SD ± 0.09) for CAR-T cells, which meets the clinical trials' release parameters. In addition, the processing of CAR-T cells formulated in CryStor yields a 91% reduction in the amount of the cryoprotectant dimethyl sulfoxide. Inertial microfluidic processing has no detrimental effects on the proliferation and cytotoxicity of CAR-T cells. Interestingly, ≈50% of T-regulatory and T-suppressor cells are depleted, suggesting the potential for inertial microfluidic processing to tune the phenotypical composition of T-cell products.

Teleconsulta en épocas de pandemia. Experiencia de los pediatras del desarrollo del Hospital de Pediatría "Prof. Dr. Juan P. Garrahan" / Teleconsultation during a pandemic. Experience of developmental pediatricians at Hospital de Pediatría "Prof. Dr. Juan P. Garrahan"

La teleconsulta es uno de los componentes de la telemedicina. Los pediatras del desarrollo realizan evaluaciones y seguimiento de niños y niñas con esafíos en su desarrollo. Durante la pandemia, debido a las restricciones de movilidad y traslado y la importancia de la continuidad de la atención, comenzaron a trabajar de manera remota realizando consultas virtuales. El objetivo del trabajo fue describir la implementación de la atención virtual, las características de los pacientes atendidos y la percepción de sus familias. Se programaron 122 teleconsultas. La media de edad de los pacientes fue de 40 ± 13 meses. Los motivos de consulta más frecuentes fueron ausencia y retraso lenguaje y dificultades conductuales. Si bien un 16 % de las familias encontró alguna dificultad, todas se mostraron agradecidas con las consultas. La atención virtual debe considerarse como un complemento de la presencialidad, que disminuye los costos y los tiempos de traslado y acerca a los pediatras del desarrollo a lugares remotos.

Teleconsultation is one of the components of telemedicine. Developmental pediatricians provide assessments and follow-up to children with developmental challenges. During the pandemic, due to movement and travel restrictions and the importance of continued care, pediatricians started working remotely with virtual consultations. The objective of this study was to describe the implementation of virtual care, the characteristics of patients seen, and their families' perception. A total of 122 teleconsultations were scheduled. Patients' mean age was 40 ± 13 months. The most common reasons for consultation were absence of and delay in language and behavioral difficulties. Although 16 % of families described some obstacles, all were grateful for the consultations. Virtual health care should be regarded as a complement to in-person care, alongside reductions in costs and travel time and the possibility of bringing developmental pediatricians to remote locations

Teleconsultation during a pandemic. Experience of developmental pediatricians at Hospital de Pediatría "Prof. Dr. Juan P. Garrahan". / Teleconsulta en épocas de pandemia. Experiencia de los pediatras del desarrollo del Hospital de Pediatría "Prof. Juan P. Garrahan".

Teleconsultation is one of the components of telemedicine. Developmental pediatricians provide assessments and followup to children with developmental challenges. During the pandemic, due to movement and travel restrictions and the importance of continued care, pediatricians started working remotely with virtual consultations. The objective of this study was to describe the implementation of virtual care, the characteristics of patients seen, and their families' perception. A total of 122 teleconsultations were scheduled. Patients' mean age was 40 ± 13 months. The most common reasons for consultation were absence of and delay in language and behavioral difficulties. Although 16 % of families described some obstacles, all were grateful for the consultations. Virtual health care should be regarded as a complement to in-person care, alongside reductions in costs and travel time and the possibility.

La teleconsulta es uno de los componentes de la telemedicina. Los pediatras del desarrollo realizan evaluaciones y seguimiento de niños y niñas con desafíos en su desarrollo. Durante la pandemia, debido a las restricciones de movilidad y traslado y la importancia de la continuidad de la atención, comenzaron a trabajar de manera remota realizando consultas virtuales. El objetivo del trabajo fue describir la implementación de la atención virtual, las características de los pacientes atendidos y la percepción de sus familias. Se programaron 122 teleconsultas. La media de edad de los pacientes fue de 40 ± 13 meses. Los motivos de consulta más frecuentes fueron ausencia y retraso lenguaje y dificultades conductuales. Si bien un 16 % de las familias encontró alguna dificultad, todas se mostraron agradecidas con las consultas. La atención virtual debe considerarse como un complemento de la presencialidad, que disminuye los costos y los tiempos de traslado y acerca a los pediatras del desarrollo a lugares remotos.

Sleep problems in a sample of children with developmental disorders, before and during the COVID-19 pandemic. / Problemas del sueño en una muestra de niñas y niños con trastornos del desarrollo, antes y durante la pandemia por COVID-19.

INTRODUCTION: Sleep problems (SPs) are common, especially among children with developmental disorders (DDs), and affect their functioning and quality of family life. Pediatricians play a major role in their management. OBJECTIVE: To define the frequency and types of SPs in a sample of children with DDs, determine the proportion of pediatricians who addressed such difficulties, assess the effects of sleep hygiene (SH), and describe the impact of the COVID-19 pandemic on sleep. POPULATION AND METHODS: This was a quasiexperiment. SPs were diagnosed based on parents' concerns and/or clinical criteria. SH strategies were provided and their effect was assessed as per parents' reports and the Children's Sleep Habits Questionnaire in Spanish (CSHQ-S) before and after the strategies. During the COVID-19 pandemic, the SP outcome measure and SH-related outcome measures were measured again. RESULTS: A total of 161 children were included. The frequency of SPs was 55 %; 80 % improved with SH. Eighty-three percent of children had a primary pediatrician; of these, 45 % had consulted about sleep. During the COVID-19 pandemic, SPs increased and SH outcome measures changed. CONCLUSION: Approximately half of children with DDs have SPs; and the problem was only addressed by 45 % of pediatricians. SH was beneficial for most children, so pediatricians' role seems critical. During the COVID-19 pandemic, SPs increased, probably as a result of its environmental impact on children with DDs.

Los problemas del sueño (PS) son frecuentes, principalmente en niñas y niños con trastornos del desarrollo (TD), y causan un impacto en su funcionamiento y calidad de vida familiar. El pediatra tiene un rol importante en su abordaje. Objetivo. Definir la frecuencia y los tipos de PS en una muestra de niñas y niños con TD, determinar la proporción de pediatras que abordaron estas dificultades, evaluar los efectos de la higiene del sueño (HS) y describir el impacto de la pandemia por COVID-19 en el sueño. Población y métodos. Estudio cuasiexperimental. El diagnóstico de PS se realizó con la preocupación de los padres y/o criterios clínicos. Se brindaron estrategias de HS, luego se evaluó su efecto según referencia de los padres y uso del cuestionario CSHQ-S (Children´s Sleep Habits Questionnaire en español) pre- y posestrategias. Durante la pandemia por COVID-19, se midió nuevamente la variable PS y las relacionadas a HS. Resultados. Se incluyeron 161 niñas y niños. La frecuencia de PS fue del 55 %. El 80 % mejoró con HS. El 83 % tenía pediatra de cabecera, y de ellos, el 45 % había preguntado acerca del sueño. Durante la pandemia por COVID-19 hubo aumento de PS y cambios en las variables de HS. Conclusión. Aproximadamente la mitad de los niñas y niños con TD presentan PS; esto solo fue abordado por el 45 % de los pediatras. La HS resultó beneficiosa para la mayoría, por lo que la intervención del pediatra parece fundamental. Durante la pandemia por COVID-19 aumentaron los PS, como posible reflejo del impacto ambiental en los niñas y niños con TD.

Problemas del sueño en una muestra de niñas y niños con trastornos del desarrollo, antes y durante la pandemia por COVID-19 / Sleep problems in a sample of children with developmental disorders, before and during the COVID-19 pandemic

Los problemas del sueño (PS) son frecuentes, principalmente en niñas y niños con trastornos del desarrollo (TD), y causan un impacto en su funcionamiento y calidad de vida familiar. El pediatra tiene un rol importante en su abordaje. Objetivo. Definir la frecuencia y los tipos de PS en una muestra de niñas y niños con TD, determinar la proporción de pediatras que abordaron estas dificultades, evaluar los efectos de la higiene del sueño (HS) y describir el impacto de la pandemia por COVID-19 en el sueño. Población y métodos. Estudio cuasiexperimental. El diagnóstico de PS se realizó con la preocupación de los padres y/o criterios clínicos. Se brindaron estrategias de HS, luego se evaluó su efecto según referencia de los padres y uso del cuestionario CSHQ-S (Children ́s Sleep Habits Questionnaire en español) pre- y posestrategias. Durante la pandemia por COVID-19, se midió nuevamente la variable PS y las relacionadas a HS. Resultados. Se incluyeron 161 niñas y niños. La frecuencia de PS fue del 55 %. El 80 % mejoró con HS. El 83 % tenía pediatra de cabecera, y de ellos, el 45 % había preguntado acerca del sueño. Durante la pandemia por COVID-19 hubo aumento de PS y cambios en las variables de HS. Conclusión. Aproximadamente la mitad de los niñas y niños con TD presentan PS; esto solo fue abordado por el 45 % de los pediatras. La HS resultó beneficiosa para la mayoría, por lo que la intervención del pediatra parece fundamental. Durante la pandemia por COVID-19 aumentaron los PS, como posible reflejo del impacto ambiental en los niñas y niños con TD.

Sleep problems (SPs) are common, especially among children with developmental disorders (DDs), and affect their functioning and quality of family life. Pediatricians play a major role in their management. Objective. To define the frequency and types of SPs in a sample of children with DDs, determine the proportion of pediatricians who addressed such difficulties, assess the effects of sleep hygiene (SH), and describe the impact of the COVID-19 pandemic on sleep. Population and methods. This was a quasi-experiment. SPs were diagnosed based on parents' concerns and/or clinical criteria. SH strategies were provided and their effect was assessed as per parents' reports and the Children's Sleep Habits Questionnaire in Spanish (CSHQ-S) before and after the strategies. During the COVID-19 pandemic, the SP outcome measure and SH-related outcome measures were measured again. Results. A total of 161 children were included. The frequency of SPs was 55 %; 80 % improved with SH. Eighty-three percent of children had a primary pediatrician; of these, 45 % had consulted about sleep. During the COVID-19 pandemic, SPs increased and SH outcome measures changed. Conclusion. Approximately half of children with DDs have SPs; and the problem was only addressed by 45 % of pediatricians. SH was beneficial for most children, so pediatricians' role seems critical. During the COVID-19 pandemic, SPs increased, probably as a result of its environmental impact on children with DD

Developing a Culturally Sensitive ICF-Based Tool to Describe Functioning of Children with Autism Spectrum Disorder: TEA-CIFunciona Version 1.0 Pilot Study.

BACKGROUND: Autism spectrum disorder (ASD) affects the daily functioning of children and their families; however, in Argentina, there are no standardized tools to guide the description, evaluation, and follow-up of functioning and disability of children with ASD. To fill this gap, the overarching purpose of this study was to create a novel tool guided by the International Classification of Functioning, Disability, and Health (ICF) Core Sets for ASD for clinical practice. METHODS: A multistep methodology was used to identify the most relevant ICF categories for an Argentinian clinical setting. The content of this ICF-based shortlist was piloted and revised according to the results. Subsequently, a toolbox of measures was proposed to operationalize each ICF category. Finally, profiles of the functioning of 100 children with ASD were created. RESULTS: An ICF-based tool called TEA-CIFunciona was created, consisting of 32 ICF categories (10 body functions, 15 activities and participation, 7 environmental factors categories). The application of TEA-CIFunciona incorporated a family-centered approach in ASD evaluations and helped identify functional needs. CONCLUSIONS: TEA-CIFunciona is the first ICF-based instrument that guides the description of functioning of children with ASD in Argentina. TEA-CIFunciona standardizes collaborative assessments in pediatric ASD populations in Latin American contexts.

Series de Pediatría Garrahan. El niño con trastornos del neurodesarrollo / Garrahan Pediatrics Series. The child with neurodevelopmental disorders

La infancia es una etapa vital clave, ya que en ella se forman los cimientos que luego determinarán el desarrollo físico y mental de las personas. Por eso, brindar las herramientas necesarias para satisfacer las necesidades y los derechos del niño es fundamental para garantizar la inclusión y la igualdad de oportunidades de todos. El desarrollo del niño conlleva cambios biológicos, psicológicos y emocionales que ocurren desde el nacimiento hasta la adolescencia, a medida que el individuo pasa de la dependencia al aumento de la autonomía. La frecuencia de los trastornos del neurodesarrollo es importante y creciente y el pediatra desempeña un papel crucial en su detección oportuna, asesoría a las familias y estrategias de manejo o derivación para asegurar las mejores posibilidades de prevención y asistencia. Asimismo, la elevada morbilidad pediátrica asociada al concepto de enfermedad crónica requiere una actitud diagnóstica anticipatoria de la disfunción neurocognitiva y madurativa a la que están expuestos estos niños. En este nuevo volumen de las Series de Pediatría Garrahan destacados profesionales estudian la temática del desarrollo desde un abordaje interdisciplinario centrado en el rol del pediatra. Entre sus características se destacan: Presentación mediante casos y ejemplos de la práctica clínica de los trastornos más frecuentes del desarrollo infantil estructurados según las diferentes etapas de lactante, preescolar y escolar, así como el abordaje de la discapacidad intelectual, el impacto de la enfermedad crónica en el aprendizaje y los marcos normativos en educación. El cierre de cada capítulo con un recordatorio de puntos clave y lecturas recomendadas, además del material complementario disponible en el sitio web. Una obra sólida y práctica, que transmite la experiencia de los profesionales del Hospital de Pediatría Prof. Dr. Juan P Garrahan, dedicada a los pediatras, dondequiera que trabajen al servicio de la salud de los niños.