Detection of minimal residual disease in acute myeloid leukemia: evaluating utility and challenges.

This study discusses the importance of minimal residual disease (MRD) detection in acute myeloid leukemia (AML) patients using liquid biopsy and next-generation sequencing (NGS). AML prognosis is based on various factors, including genetic alterations. NGS has revealed the molecular complexity of AML and helped refine risk stratification and personalized therapies. The long-term survival rates for AML patients are low, and MRD assessment is crucial in predicting prognosis. Currently, the most common methods for MRD detection are flow cytometry and quantitative PCR, but NGS is being incorporated into clinical practice due to its ability to detect genomic aberrations in the majority of AML patients. Typically, bone marrow samples are used for MRD assessment, but using peripheral blood samples or liquid biopsies would be less invasive. Leukemia originates in the bone marrow, along with the cfDNA obtained from peripheral blood. This study aimed to assess the utility of cell-free DNA (cfDNA) from peripheral blood samples for MRD detection in AML patients. A cohort of 20 AML patients was analyzed using NGS, and a correlation between MRD assessment by cfDNA and circulating tumor cells (CTCs) in paired samples was observed. Furthermore, a higher tumor signal was detected in cfDNA compared to CTCs, indicating greater sensitivity. Challenges for the application of liquid biopsy in MRD assessment were discussed, including the selection of appropriate markers and the sensitivity of certain markers. This study emphasizes the potential of liquid biopsy using cfDNA for MRD detection in AML patients and highlights the need for further research in this area.

Corrigendum: Personalized monitoring of circulating tumor DNA with a specific signature of trackable mutations after chimeric antigen receptor T-cell therapy in follicular lymphoma patients.

[This corrects the article DOI: 10.3389/fimmu.2023.1188818.].

Personalized monitoring of circulating tumor DNA with a specific signature of trackable mutations after chimeric antigen receptor T-cell therapy in follicular lymphoma patients.

Background: CART therapy has produced a paradigm shift in the treatment of relapsing FL patients. Strategies to optimize disease surveillance after these therapies are increasingly necessary. This study explores the potential value of ctDNA monitoring with an innovative signature of personalized trackable mutations. Method: Eleven FL patients treated with anti-CD19 CAR T-cell therapy were included. One did not respond and was excluded. Genomic profiling was performed before starting lymphodepleting chemotherapy to identify somatic mutations suitable for LiqBio-MRD monitoring. The dynamics of the baseline mutations (4.5 per patient) were further analyzed on 59 cfDNA follow-up samples. PET/CT examinations were performed on days +90, +180, +365, and every six months until disease progression or death. Results: After a median follow-up of 36 months, all patients achieved a CR as the best response. Two patients progressed. The most frequently mutated genes were CREBBP, KMT2D and EP300. Simultaneous analysis of ctDNA and PET/CT was available for 18 time-points. When PET/CT was positive, two out of four ctDNA samples were LiqBio-MRD negative. These two negative samples corresponded to women with a unique mesenteric mass in two evaluations and never relapsed. Meanwhile, 14 PET/CT negative images were mutation-free based on our LiqBio-MRD analysis (100%). None of the patients had a negative LiqBio-MRD test by day +7. Interestingly, all durably responding patients had undetectable ctDNA at or around three months after infusion. Two patients presented discordant results by PET/CT and ctDNA levels. No progression was confirmed in these cases. All the progressing patients were LiqBio-MRD positive before progression. Conclusion: This is a proof-of-principle for using ctDNA to monitor response to CAR T-cell therapy in FL. Our results confirm that a non-invasive liquid biopsy MRD analysis may correlate with response and could be used to monitor response. Harmonized definitions of ctDNA molecular response and pinpointing the optimal timing for assessing ctDNA responses are necessary for this setting. If using ctDNA analysis, we suggest restricting follow-up PET/CT in CR patients to a clinical suspicion of relapse, to avoid false-positive results.

Real-life disease monitoring in follicular lymphoma patients using liquid biopsy ultra-deep sequencing and PET/CT.

In the present study, we screened 84 Follicular Lymphoma patients for somatic mutations suitable as liquid biopsy MRD biomarkers using a targeted next-generation sequencing (NGS) panel. We found trackable mutations in 95% of the lymph node samples and 80% of the liquid biopsy baseline samples. Then, we used an ultra-deep sequencing approach with 2 · 10-4 sensitivity (LiqBio-MRD) to track those mutations on 151 follow-up liquid biopsy samples from 54 treated patients. Positive LiqBio-MRD at first-line therapy correlated with a higher risk of progression both at the interim evaluation (HRINT 11.0, 95% CI 2.10-57.7, p = 0.005) and at the end of treatment (HREOT, HR 19.1, 95% CI 4.10-89.4, p < 0.001). Similar results were observed by PET/CT Deauville score, with a median PFS of 19 months vs. NR (p < 0.001) at the interim and 13 months vs. NR (p < 0.001) at EOT. LiqBio-MRD and PET/CT combined identified the patients that progressed in less than two years with 88% sensitivity and 100% specificity. Our results demonstrate that LiqBio-MRD is a robust and non-invasive approach, complementary to metabolic imaging, for identifying FL patients at high risk of failure during the treatment and should be considered in future response-adapted clinical trials.

Corrigendum: Personalized monitoring of circulating tumor DNA with a specific signature of trackable mutations after chimeric antigen receptor T-cell therapy in follicular lymphoma patients.

[This corrects the article DOI: 10.3389/fimmu.2023.1188818.].

Detection of kinase domain mutations in BCR::ABL1 leukemia by ultra-deep sequencing of genomic DNA.

The screening of the BCR::ABL1 kinase domain (KD) mutation has become a routine analysis in case of warning/failure for chronic myeloid leukemia (CML) and B-cell precursor acute lymphoblastic leukemia (ALL) Philadelphia (Ph)-positive patients. In this study, we present a novel DNA-based next-generation sequencing (NGS) methodology for KD ABL1 mutation detection and monitoring with a 1.0E-4 sensitivity. This approach was validated with a well-stablished RNA-based nested NGS method. The correlation of both techniques for the quantification of ABL1 mutations was high (Pearson r = 0.858, p < 0.001), offering DNA-DeepNGS a sensitivity of 92% and specificity of 82%. The clinical impact was studied in a cohort of 129 patients (n = 67 for CML and n = 62 for B-ALL patients). A total of 162 samples (n = 86 CML and n = 76 B-ALL) were studied. Of them, 27 out of 86 harbored mutations (6 in warning and 21 in failure) for CML, and 13 out of 76 (2 diagnostic and 11 relapse samples) did in B-ALL patients. In addition, in four cases were detected mutation despite BCR::ABL1 < 1%. In conclusion, we were able to detect KD ABL1 mutations with a 1.0E-4 sensitivity by NGS using DNA as starting material even in patients with low levels of disease.

Serum levels of inflammatory mediators as prognostic biomarker in silica exposed workers.

Silicosis is a diffuse interstitial lung disease caused by sustained inhalation of silica and silicates. Several cytokines are activated by their inhalation and can mediate the process of pulmonary fibrosis. The identification of biomarkers could allow an early diagnosis before the development of radiological alterations and help monitor the evolution of patients. The objetive of this study was to determine the clinical significance of specific biomarkers, to estimate their association with the development, severity and/or progression of silicosis, and identify determinants of this evolution. We conducted a prospective observational study in patients attending the pulmonology clinic from 2009 to 2018. Serum levels of the following inflammatory mediators were assessed: interleukin-6 (IL-6), interleukin 2 receptor subunit alpha (IL2R) interleukin 1 beta (IL1B), interleukin-8 (IL-8), tumour necrosis factor-alpha (TNF-α), transforming growth factor-beta1 (TGF-ß1), alpha-1 antitrypsin (AAT), C-reactive protein (CRP), lactate dehydrogenase (LDH) and ferritin in subjects exposed to silica, with and without silicosis. Association between those inflammatory mediators with lung function measurements and radiological severity of disease and their impact on prognosis were analysed. 337 exposed to silica (278 with silicosis) and 30 subjects in the control group were included. IL-8, α1AT, ferritin, CRP and LDH levels were higher in silicosis than in those exposed to silica without silicosis. IL-8, LDH and AAT levels were associated with progression of silicosis and IL-6, IL-8, LDH, AAT, ferritin, and CRP with vital status. The results of the ROC analysis indicated the potential of IL-8 as a biomarker in the presence of silicosis and for the prediction of mortality.

Mapping the clinical pharmacokinetics and pharmacogenetic units operating in Spanish hospitals.

OBJECTIVE: The goal of this article is to analyze the situation of  harmacokinetics and pharmacogenetics units in the pharmacy departments of  Spanish hospitals, evaluate their development both in the clinical and educational areas, and draw up a map reflecting their current status.  Method: A 29-item survey structured in five blocks was designed with general questions about the respondents' hospital and the clinical and educational activities carried out by their pharmacy department, in the fields of both pharmacokinetics and pharmacogenetics. RESULTS: Sixty-nine  hospitals answered the survey. The highest response rates corresponded to  Catalonia, the Valencia region and Andalusia. The drug families subject to closest monitoring were classic antibiotics (93%), digoxin (57%), classic antiepileptics  (51%) and biologicals (43%). The most frequently used computer programs  included PKS and NONMEM (93% and 22% of hospitals, respectively).  Regarding training in pharmacokinetics, second year residents were those who  most frequently rotated through the pharmacokinetics unit (40%), while 44%  of those units allowed external residents. As far as pharmacogenetics is  concerned, in 42% of hospitals that engaged in pharmacogenetic work, the  department in charge was pharmacy. The most frequent specialties covered  were hemato-oncology (72%) and psychiatry (15%). Twenty-four percent of  hospitals offered rotations through their pharmacogenetics unit but only seven of them allowed external residents. CONCLUSIONS: The results of the survey showed an increase in the  erformance of pharmacokinetic and pharmacogenetic activities by Spanish hospital pharmacies as compared with the data from a 2009 baseline  survey, with many hospitals introducing the performance of therapeutic drug monitoring of non-classical antibiotics, immunosuppressants and biologics. However, the percentage of hospitals that follow the ideal model based on analytical determinations and pharmacokinetic reporting has decreased the data obtained served as a basis to create an updated map of the  harmacokinetics and pharmacogenetics units operating in Spanish hospital  pharmacy departments. This map, available at http://bit.ly/mapaPKGen, will be very useful to facilitate the training of residents in these disciplines and will  help promote the development of pharmacokinetic and pharmacogenetic activities among hospital pharmacists.

OBJETIVO: Dar a conocer la actividad asistencial y docente de las unidades de  farmacocinética y farmacogenética de los servicios de farmacia hospitalaria  españoles y elaborar un mapa que refleje la situación actual. Método: Se  diseñó una encuesta de 29 preguntas estructuradas en cinco bloques: datos  enerales del hospital e información sobre la actividad asistencial y docente,  tanto en el área de farmacocinética como de farmacogenética en los servicios  de farmacia hospitalaria. RESULTADOS: Respondieron la encuesta 69 hospitales. Las regiones geográficas  con mayor número de respuestas fueron Cataluña, Comunidad   Valenciana y Andalucía. Los grupos farmacológicos que más se monitorizaron fueron los antibióticos clásicos (vancomicina y aminoglucósidos)  (93%), digoxina (57%), antiepilépticos clásicos (51%) y biológicos (43%). Los  programas informáticos que con más frecuencia se utilizaban fueron PKS y NONMEM, con un 93% y 22%, respectivamente. Respecto a la docencia en  farmacocinética, fue el segundo año de residencia cuando la mayoría de los farmacéuticos internos residentes rotaban por el área (40%) y un 44% de las  unidades permitían rotantes externos. El responsable de la farmacogenética era el servicio de farmacia hospitalario en un 43% de los  casos. Los ámbitos más frecuentes fueron oncohematología (72%) y  psiquiatría (15%). Un 24% de los hospitales ofrecían rotación por la unidad de  farmacogenética y sólo 7 servicios de farmacia ofertaron rotaciones externas. Conclusiones: Los resultados de la encuesta mostraron un incremento en la  realización de actividades de farmacocinética y farmacogenética en los servicios de farmacia hospitalaria comparados con los datos de la encuesta  basal de 2009. Se inició la realización de monitorización terapéutica de biológicos, inmunosupresores y antibióticos no clásicos. Sin embargo, ha disminuido el porcentaje de hospitales que aplican el modelo ideal basado en la  realización de determinación analítica e informe farmacocinético desde  farmacia. Los datos obtenidos permiten disponer de un mapa actualizado de  las unidades de farmacocinética y farmacogenética clínicas en los servicios de  farmacia hospitalaria españoles. Esta información se encuentra disponible en  http://bit.ly/mapaPKGen y será de gran utilidad para facilitar la formación de  nuestros residentes en estas disciplinas y ayudará a promocionar su desarrollo  entre los farmacéuticos de hospital.

Mapa de unidades de farmacocinética y farmacogenética clínica en España / Mapping the clinical pharmacokinetics and pharmacogenetic units operating in Spanish hospitals

Objetivo: Dar a conocer la actividad asistencial y docente de las unidades de farmacocinética y farmacogenética de los servicios de farmaciahospitalaria españoles y elaborar un mapa que refleje la situación actual.Método: Se diseñó una encuesta de 29 preguntas estructuradas encinco bloques: datos generales del hospital e información sobre la actividad asistencial y docente, tanto en el área de farmacocinética como defarmacogenética en los servicios de farmacia hospitalaria.Resultados: Respondieron la encuesta 69 hospitales. Las regiones geográficas con mayor número de respuestas fueron Cataluña, ComunidadValenciana y Andalucía. Los grupos farmacológicos que más se monitorizaron fueron los antibióticos clásicos (vancomicina y aminoglucósidos) (93%),digoxina (57%), antiepilépticos clásicos (51%) y biológicos (43%). Los programas informáticos que con más frecuencia se utilizaban fueron PKS yNONMEM, con un 93% y 22%, respectivamente. Respecto a la docenciaen farmacocinética, fue el segundo año de residencia cuando la mayoría delos farmacéuticos internos residentes rotaban por el área (40%) y un 44%de las unidades permitían rotantes externos. El responsable de la farmacogenética era el servicio de farmacia hospitalario en un 43% de los casos.Los ámbitos más frecuentes fueron oncohematología (72%) y psiquiatría(15%). Un 24% de los hospitales ofrecían rotación por la unidad de farmacogenética y sólo 7 servicios de farmacia ofertaron rotaciones externas. Conclusiones: Los resultados de la encuesta mostraron un incremento enla realización de actividades de farmacocinética y farmacogenética en losservicios de farmacia hospitalaria comparados con los datos de la encuesta basal de 2009. (AU)

Objective: The goal of this article is to analyze the situation of pharmacokinetics and pharmacogenetics units in the pharmacy departmentsof Spanish hospitals, evaluate their development both in the clinical andeducational areas, and draw up a map reflecting their current status.Method: A 29-item survey structured in five blocks was designed withgeneral questions about the respondents’ hospital and the clinical andeducational activities carried out by their pharmacy department, in thefields of both pharmacokinetics and pharmacogenetics.Results: Sixty-nine hospitals answered the survey. The highest response ratescorresponded to Catalonia, the Valencia region and Andalusia. The drugfamilies subject to closest monitoring were classic antibiotics (93%), digoxin(57%), classic antiepileptics (51%) and biologicals (43%). The most frequentlyused computer programs included PKS and NONMEM (93% and 22% ofhospitals, respectively). Regarding training in pharmacokinetics, second yearresidents were those who most frequently rotated through the pharmacokinetics unit (40%), while 44% of those units allowed external residents. Asfar as pharmacogenetics is concerned, in 42% of hospitals that engaged inpharmacogenetic work, the department in charge was pharmacy. The mostfrequent specialties covered were hemato-oncology (72%) and psychiatry(15%). Twenty-four percent of hospitals offered rotations through their pharmacogenetics unit but only seven of them allowed external residents.Conclusions: The results of the survey showed an increase in the performance of pharmacokinetic and pharmacogenetic activities by Spanishhospital pharmacies as compared with the data from a 2009 baseline survey, with many hospitals introducing the performance of therapeutic drugmonitoring of non-classical antibiotics, immunosuppressants and biologics. (AU)

A multicentre prospective study evaluating the impact of proton-pump inhibitors omeprazole and pantoprazole on voriconazole plasma concentrations.

Voriconazole is an antifungal metabolised by CYP2C19 enzyme, which can be inhibited by proton-pump inhibitors (PPIs). A prospective observational study was carried out to determine the influence of PPIs on voriconazole pharmacokinetic. The 78 patients included were divided into 4 groups: omeprazole (n = 32), pantoprazole (n = 25), esomeprazole (n = 3) and no PPI (n = 18). Patients with no PPI had no significant difference in plasma voriconazole concentration when compared with those with PPI (2.63 ± 2.13 µg/mL [95% confidence interval {CI} 1.57-3.69] vs 2.11 ± 1.73 µg/mL [95%CI 1.67-2.55], P > .05). However, voriconazole plasma concentration was significantly lower in patients treated with pantoprazole vs those treated with omeprazole (1.44 ± 1.22 µg/mL [95%CI 0.94-1.94) vs 2.67 ± 1.88 µg/mL [95%CI 2.02-3.32], P = .013) suggesting a greater CYP2C19 enzyme inhibitory effect of omeprazole. This study demonstrates the greater CYP inhibition capacity of omeprazole and should be helpful for the choice of PPIs for patients treated with voriconazole.

Impact of CYP2C19 Genotype and Drug Interactions on Voriconazole Plasma Concentrations: A Spain Pharmacogenetic-Pharmacokinetic Prospective Multicenter Study.

BACKGROUND: Voriconazole, a first-line agent for the treatment of invasive fungal infections, is mainly metabolized by cytochrome P450 (CYP) 2C19. A significant portion of patients fail to achieve therapeutic voriconazole trough concentrations, with a consequently increased risk of therapeutic failure. OBJECTIVE: To show the association between subtherapeutic voriconazole concentrations and factors affecting voriconazole pharmacokinetics: CYP2C19 genotype and drug-drug interactions. METHODS: Adults receiving voriconazole for antifungal treatment or prophylaxis were included in a multicenter prospective study conducted in Spain. The prevalence of subtherapeutic voriconazole troughs was analyzed in the rapid metabolizer and ultra-rapid metabolizer patients (RMs and UMs, respectively), and compared with the rest of the patients. The relationship between voriconazole concentration, CYP2C19 phenotype, adverse events (AEs), and drug-drug interactions was also assessed. RESULTS: In this study 78 patients were included with a wide variability in voriconazole plasma levels with only 44.8% of patients attaining trough concentrations within the therapeutic range of 1 and 5.5 µg/ml. The allele frequency of *17 variant was found to be 29.5%. Compared with patients with other phenotypes, RMs and UMs had a lower voriconazole plasma concentration (RM/UM: 1.85 ± 0.24 µg/ml vs other phenotypes: 2.36 ± 0.26 µg/ml). Adverse events were more common in patients with higher voriconazole concentrations (p<0.05). No association between voriconazole trough concentration and other factors (age, weight, route of administration, and concomitant administration of enzyme inducer, enzyme inhibitor, glucocorticoids, or proton pump inhibitors) was found. CONCLUSION: These results suggest the potential clinical utility of using CYP2C19 genotype-guided voriconazole dosing to achieve concentrations in the therapeutic range in the early course of therapy. Larger studies are needed to confirm the impact of pharmacogenetics on voriconazole pharmacokinetics.

An Observational Study of the Efficacy and Safety of Voriconazole in a Real-Life Clinical Setting.

An observational retrospective study in patients treated with voriconazole was made to evaluate outcomes, safety, drug interactions and characteristics of the treatment. A total of 96 patients were included. In 78.12%, at least one inducer or enzyme inhibitor was detected. The most frequently observed potential interaction was the simultaneous administration of omeprazole. A large number of patients were concurrently treated with corticosteroids. The simultaneous administration of drugs acting as CYP450 enzyme inhibitors was associated with a higher risk of toxicity while concomitant administration of corticosteroids seemed a protective factor. Our study is one of the few ones, which evaluate the use of voriconazole in a real life clinical setting. We demonstrate the wide variety of strategies in the voriconazole using and the large number of dugs that are susceptible to pharmacokinetic interactions. This study reinforces the need to implement voriconazole pharmacokinetic monitoring in order to optimize antifungal treatment.

Ophthalmic Econazole Hydrogels for the Treatment of Fungal Keratitis.

Econazole is a feasible alternative treatment in the management of fungal keratitis. Nevertheless, its low water solubility is considered the main limitation to the incorporation into ophthalmic formulations. In this work, econazole nitrate is solubilized by using cyclodextrins to achieve an optimum therapeutic concentration. Phase solubility diagrams suggest α-cyclodextrin as the most effective cyclodextrin and later the inclusion complex formed with this one was characterized in solution by 1D, 2D-NMR, and molecular modeling. Econazole-α-cyclodextrin inclusion complex was included in 2 types of ocular hydrogels: a natural polysaccharides ion-sensitive hydrogel and a hyaluronic acid hydrogel. Both of them show no ocular irritation in the hen's egg test on chorioallantoic membrane assay and a controlled econazole release over time. Permeability studies suggest that hydrogels do not modify the econazole nitrate permeability through bovine cornea in comparison with an econazole-α-cyclodextrin inclusion complex solution. Finally, ocular biopermanence studies performed using positron emission tomography show these hydrogels present a high retention time on the eye. Results suggest the developed formulations have a high potential as vehicles for the econazole topical ocular administration as fungal keratitis treatment.

61.º Congreso de la SEFH. Nutrición parenteral libre de aminoácidos ramificados para el manejo urgente de una descompensación grave de la enfermedad del jarabe de arce / No disponible

No disponible

61.º Congreso de la SEFH. Complicaciones asociadas con medicamentos en un postoperatorio tórpido de trasplante hepático / No disponible

No disponible

Progression of liver stiffness predicts clinical events in HIV/HCV-coinfected patients with compensated cirrhosis.

BACKGROUND: Our objective was to assess the predictive value of the changes of liver stiffness (LS) for clinical outcome in HIV/HCV-coinfected patients with compensated liver cirrhosis and a LS value < 40 kPa. METHODS: Prospective cohort of 275 HIV/HCV-coinfected patients with cirrhosis, no previous liver decompensation (LD) and LS < 40 kPa. The time from diagnosis to LD and/or hepatocellular carcinoma (HCC) and the predictors of this outcome were evaluated. Significant progression of LS was defined as an increase ≥ 30 % over the baseline value at any time during the follow-up. RESULTS: After a median (Q1-Q3) follow-up of 32 (20-48) months, 19 (6.9 %, 95 % CI: 3.8 %-9.9 %) patients developed a first LD and/or HCC. At the end of the follow-up, 247 (90 %) patients had undergone a further LS examination. Of them, 77 (31 %) patients had a significant progression of LS. The mean (SD) survival time free of LD and/or HCC was 67 (3) and 77 (1) months in patients with or without significant progression of LS (p = 0.01). Significant progression of LS was an independent predictor of LD and/or HCC (Adjusted Hazard Ratio 4.63; 95 % confidence interval: 1.34-16.02; p = 0.015). CONCLUSIONS: Significant progression of LS is associated with a higher risk of clinical events in HIV/HCV-coinfected patients with compensated cirrhosis and LS < 40 kPa.