High cell-free DNA is associated with disease progression, inflammasome activation and elevated levels of inflammasome-related cytokine IL-18 in patients with myelofibrosis

Myelofibrosis (MF) is a clonal hematopoietic stem cell disorder classified among chronic myeloproliferative neoplasms, characterized by exacerbated myeloid and megakaryocytic proliferation and bone marrow fibrosis. It is induced by driver (JAK2/CALR/MPL) and high molecular risk mutations coupled to a sustained inflammatory state that contributes to disease pathogenesis. Patient outcome is determined by stratification into risk groups and refinement of current prognostic systems may help individualize treatment decisions. Circulating cell-free (cf)DNA comprises short fragments of double-stranded DNA, which promotes inflammation by stimulating several pathways, including inflammasome activation, which is responsible for IL-1β and IL-18 maturation and release. In this work, we assessed the contribution of cfDNA as a marker of disease progression and mediator of inflammation in MF. cfDNA was increased in MF patients and higher levels were associated with adverse clinical outcome, a high-risk molecular profile, advanced disease stages and inferior overall survival, indicating its potential value as a prognostic marker. Cell-free DNA levels correlated with tumor burden parameters and markers of systemic inflammation. To mimic the effects of cfDNA, monocytes were stimulated with poly(dA:dT), a synthetic double-stranded DNA. Following stimulation, patient monocytes released higher amounts of inflammasome-processed cytokine, IL-18 to the culture supernatant, reflecting enhanced inflammasome function. Despite overexpression of cytosolic DNA inflammasome sensor AIM2, IL-18 release from MF monocytes was shown to rely mainly on the NLRP3 inflammasome, as it was prevented by NLRP3-specific inhibitor MCC950. Circulating IL-18 levels were increased in MF plasma, reflecting in vivo inflammasome activation, and highlighting the previously unrecognized involvement of this cytokine in MF cytokine network. Monocyte counts were higher in patients and showed a trend towards correlation with IL-18 levels, suggesting monocytes represent a source of circulating IL-18. The close correlation shown between IL-18 and cfDNA levels, together with the finding of enhanced DNA-triggered IL-18 release from monocytes, suggest that cfDNA promotes inflammation, at least in part, through inflammasome activation. This work highlights cfDNA, the inflammasome and IL-18 as additional players in the complex inflammatory circuit that fosters MF progression, potentially providing new therapeutic targets.Fil: de Luca, Geraldine. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Lev, Paola Roxana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Camacho, Maria F.. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaFil: Goette, Nora Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Sackmann, Federico. Fundación Para Combatir la Leucemia; ArgentinaFil: Castro Ríos, Miguel A.. No especifíca;Fil: Moiraghi, Beatriz. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos "Ramos Mejía"; ArgentinaFil: Cortes Guerrieri, Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Bendek, Georgina. Hospital Italiano; ArgentinaFil: Carricondo, Emiliano. Universidad Austral. Hospital Universitario Austral; ArgentinaFil: Enrico, Alicia. Hospital Italiano; ArgentinaFil: Vallejo, Veronica. Instituto Cardiovascular de Buenos Aires; ArgentinaFil: Varela, Ana. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos "Ramos Mejía"; ArgentinaFil: Khoury, Marina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Gutierrez, Marina. Laboratorio Stamboulian; ArgentinaFil: Larripa, Irene Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaFil: Marta, Rosana Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaFil: Glembotsky, Ana Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Heller, Paula Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentin

High cell-free DNA is associated with disease progression, inflammasome activation and elevated levels of inflammasome-related cytokine IL-18 in patients with myelofibrosis

Myelofibrosis (MF) is a clonal hematopoietic stem cell disorder classified among chronic myeloproliferative neoplasms, characterized by exacerbated myeloid and megakaryocytic proliferation and bone marrow fibrosis. It is induced by driver (JAK2/CALR/MPL) and high molecular risk mutations coupled to a sustained inflammatory state that contributes to disease pathogenesis. Patient outcome is determined by stratification into risk groups and refinement of current prognostic systems may help individualize treatment decisions. Circulating cell-free (cf)DNA comprises short fragments of double-stranded DNA, which promotes inflammation by stimulating several pathways, including inflammasome activation, which is responsible for IL-1β and IL-18 maturation and release. In this work, we assessed the contribution of cfDNA as a marker of disease progression and mediator of inflammation in MF. cfDNA was increased in MF patients and higher levels were associated with adverse clinical outcome, a high-risk molecular profile, advanced disease stages and inferior overall survival, indicating its potential value as a prognostic marker. Cell-free DNA levels correlated with tumor burden parameters and markers of systemic inflammation. To mimic the effects of cfDNA, monocytes were stimulated with poly(dA:dT), a synthetic double-stranded DNA. Following stimulation, patient monocytes released higher amounts of inflammasome-processed cytokine, IL-18 to the culture supernatant, reflecting enhanced inflammasome function. Despite overexpression of cytosolic DNA inflammasome sensor AIM2, IL-18 release from MF monocytes was shown to rely mainly on the NLRP3 inflammasome, as it was prevented by NLRP3-specific inhibitor MCC950. Circulating IL-18 levels were increased in MF plasma, reflecting in vivo inflammasome activation, and highlighting the previously unrecognized involvement of this cytokine in MF cytokine network. Monocyte counts were higher in patients and showed a trend towards correlation with IL-18 levels, suggesting monocytes represent a source of circulating IL-18. The close correlation shown between IL-18 and cfDNA levels, together with the finding of enhanced DNA-triggered IL-18 release from monocytes, suggest that cfDNA promotes inflammation, at least in part, through inflammasome activation. This work highlights cfDNA, the inflammasome and IL-18 as additional players in the complex inflammatory circuit that fosters MF progression, potentially providing new therapeutic targets

Consensus guidelines for the use and interpretation of angiogenesis assays

The formation of new blood vessels, or angiogenesis, is a complex process that plays important roles in growth and development, tissue and organ regeneration, as well as numerous pathological conditions. Angiogenesis undergoes multiple discrete steps that can be individually evaluated and quantified by a large number of bioassays. These independent assessments hold advantages but also have limitations. This article describes in vivo, ex vivo, and in vitro bioassays that are available for the evaluation of angiogenesis and highlights critical aspects that are relevant for their execution and proper interpretation. As such, this collaborative work is the first edition of consensus guidelines on angiogenesis bioassays to serve for current and future reference

Moxetumomab pasudotox. Inmunotoxina recombinante anti-CD22

Hairy cell leukemia (HCL) is a rare and incurable disease that represents approximately 2% of lymphoid leukemias. Treatment is indicated for patients with symptomatic disease manifested by splenomegaly, cytopenias or serious infections. Purine nucleoside analogs (PNA), such as cladribine and pentostatin, are the preferred drugs for first line treatment in most cases. With PNA high rates of complete response and long disease-free periods are obtained, however, relapses are frequent. Second line treatment includes PNA combined with rituximab.Even active, recurring PNA are associated with a significant accumulated toxicity. Hence, the need for investigate new therapeutic alternatives as moxetumomab pasudotox (CAT-8015). Two multicenter studies, one in phase I and one in phase III, had positive results, since moxetumomab pasudotox (CAT-8015) achieved high rates of complete responses, lasting responses and negative minimal residual disease, also proving safe for patients with relapsed or refractory HCL.La leucemia de células vellosas (LCV) es una enfermedadrara e incurable que representa aproximadamente el 2% de las leucemias linfoides. El tratamiento está indicado en pacientes con enfermedad sintomática, ya sea por esplenomegalia, citopenias o infecciones graves. Los análogos de nucleósidos de purinas (ANP), como cladribina y pentostatina, son las drogas de elección para el tratamiento de primera línea en la mayoría de los casos, logrando altas tasas de remisión y largos períodos libres de enfermedad. Sin embargo, las recaídas son frecuentes.Los tratamientos de segunda línea incluyen ANP combinados con rituximab pero, si bien conservan eficacia, las terapias recurrentes se asocian a una toxicidad acumulada importante. Por tal motivo, es de suma importancia investigar y desarrollar nuevas alternativas terapéuticas como moxetumomab pasudotox (CAT-8015).Dos estudios multicéntricos, uno en fase I y otro en fase III, tuvieron resultados positivos, ya que moxetumomab pasudotox (CAT-8015) logró tasas elevadas de respuestas completas, respuestas duraderas y enfermedad mínima residual negativa demostrando, además, ser seguro para pacientes con LCV recaídos o refractarios

Tratamiento de la leucemia linfocítica aguda (LLA) en niños

Artículo científico -- Universidad de Costa Rica, Instituto de Investigaciones en Salud. 1985Se reportan los resultados del tratamiento antileucémico en 195 niños con LLA. Noventa y tres porciento de los pacientes hicieron remisión completa, con 5% de mortalidad durante Ia inducción El 81% de los niños permanecen en remisión completa continuada, siendo Ia duración de Ia remisión significativamente menor en los pacientes de mal pronóstico. El sitio más frecuente de recaída fue la médula ósea (11%). Se revisaron las complicaciones durante Ia inducción, y se concluye que con buena terapia de apoyo es factible realizar el protocolo BFM en países de América Latina, con resultados similares a los reportados por otros investigadores. Este estudio ha sido realizado bajo el Programa Colaborativo de Investigación en el Tratamiento de Cáncer, un proyecto auspiciado por la Organizacien Panamericana de Ia Salud y el Instituto Nacional de Cáncer de los Estados Unidos de Norteamérica bajo contrato N° NO1-CO-27391. Se realizó en los siguientes centros por los investigadores que se citan: 1. Hospital de Ninos (Buenos Aires): F. Sackmann, J. Peñalver, J. Braier, V. Maro, P. Bustelo. 2. Hospital Nacional de Niños (Costa Rica); E. Jiménez, J.M. Carrillo, M. Navarrete, E. Quesada, C. Odio, F. Lobo, R. Jiménez, L. Mora, I. García. 3. Cuba: E. Svarch, M. Lagarde, B. Vergara, J. Méndez. 4. Instituto de Investigaciones Hematológicas (Buenos Aires): M. Tezanos, G. Garay, J. Dupont, S. Bruno, S. Pavlovsky, C. Scaglione, N. Roizman, N. Fismann, 5. Instituto Municipal de Hematología (Buenos Aires): M. Eppinger-Helft, G. Hidalgo.Universidad de Costa Rica, Instituto de Investigaciones en SaludUCR::Investigación::Unidades de Investigación::Ciencias de la Salud::Instituto de Investigaciones en Salud (INISA

Impact of Ibrutinib in Quality of Life (QoL) in Patients with Chronic Lymphocytic Leukemia (CLL): Preliminary Results of Real-World Experience

Context: Clinical data from controlled trials report an improvement in QoL in patients with CLL in ibrutinib monotherapy. Reallife evidence is necessary to confirm this data. Objective: The disease control when treatment is initiated could lead to decreasedfatigue. It is likely that treatment with ibrutinib could result in animprovement in QoL in our real-world population. Primaryobjective: evaluate impact of ibrutinib treatment in QoL. Wedefined a clinically meaningful improvement 3 points in FACITfatigue score. Secondary Endpoints: Detect a 10% improvement byEQ5D VAS. Correlate baseline and follow-up hemoglobin levels(meaningful improvement 1g/dL). Design: This is a prospective,longitudinal, single arm study enrolling consecutive CLL patientsunder ibrutinib monotherapy either as first or further line oftreatment. Median follow-up was 7 months (range 1-28). Setting:Patients are recruited in an academic referral center in BuenosAires. Interventions: QoL was explored with FACIT-fatigue andEQ5D visual-analogue-scale (VAS) questionnaires (with copyrightpermission). Assessment by results reported by patient questionnaires are completed on months 0-1-3-6 and 12 since the beginningof treatment. Main Outcomes Measures: We are reporting preliminary results after 3 months of treatment compared to baseline.Statistical Analysis: data was analyzed with the Sign Test (BinomialTest). Results: A total of 21 CLL patients who started ibrutinibbetween 2016 and 2018 were included. Median age was 75 years(range 57-84); 12 patients (57%) were males. Ibrutinib was first-linetherapy in 7 patients (33%), second-line in 7 patients (33%) and 7patients (33%) received 3 previous lines. After 3 months oftreatment, the median change in FACIT-fatigue score 3 pointswas reached in 13 patients (62%) as compared to baseline(p=0.024). After 3 months of treatment there was no evidence of amedian change >10% on the EQ5D VAS (p=0.593) nor a significant improvement on hemoglobin level (p=0.105).Conclusions: These results suggest an early improvement of fatiguewithin the first 3 months of treatment with ibrutinib. Longerfollow-up and larger number of patient are necessary to confirm thisdata and determine the further improvements of QoL withcontinuous treatment and correlation with hemoglobin changes.Fil: Mela Osorio, Maria Jose. Fundación Para Combatir la Leucemia; ArgentinaFil: Pavlovsky, Carolina. Fundación Para Combatir la Leucemia; ArgentinaFil: Pavlovsky, Astrid. Fundación Para Combatir la Leucemia; ArgentinaFil: Fernandez, Isolda. Fundación Para Combatir la Leucemia; ArgentinaFil: Massa, Federico Sackmann. Fundación Para Combatir la Leucemia; ArgentinaFil: Ferrari, Luciana. Fundación Para Combatir la Leucemia; ArgentinaFil: Juni, Mariana. Fundación Para Combatir la Leucemia; ArgentinaFil: Riddick, Maximiliano Luis. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Matemáticas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Cálculo; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Pavlovsky, Miguel A.. Fundación Para Combatir la Leucemia; Argentin