Next-generation BCMA-targeted chimeric antigen receptor CARTemis-1: the impact of manufacturing procedure on CAR T-cell features.

PURPOSE: CAR therapy targeting BCMA is under investigation as treatment for multiple myeloma. However, given the lack of plateau in most studies, pursuing more effective alternatives is imperative. We present the preclinical and clinical validation of a new optimized anti-BCMA CAR (CARTemis-1). In addition, we explored how the manufacturing process could impact CAR-T cell product quality and fitness. METHODS: CARTemis-1 optimizations were evaluated at the preclinical level both, in vitro and in vivo. CARTemis-1 generation was validated under GMP conditions, studying the dynamics of the immunophenotype from leukapheresis to final product. Here, we studied the impact of the manufacturing process on CAR-T cells to define optimal cell culture protocol and expansion time to increase product fitness. RESULTS: Two different versions of CARTemis-1 with different spacers were compared. The longer version showed increased cytotoxicity. The incorporation of the safety-gene EGFRt into the CARTemis-1 structure can be used as a monitoring marker. CARTemis-1 showed no inhibition by soluble BCMA and presents potent antitumor effects both in vitro and in vivo. Expansion with IL-2 or IL-7/IL-15 was compared, revealing greater proliferation, less differentiation, and less exhaustion with IL-7/IL-15. Three consecutive batches of CARTemis-1 were produced under GMP guidelines meeting all the required specifications. CARTemis-1 cells manufactured under GMP conditions showed increased memory subpopulations, reduced exhaustion markers and selective antitumor efficacy against MM cell lines and primary myeloma cells. The optimal release time points for obtaining the best fit product were > 6 and < 10 days (days 8-10). CONCLUSIONS: CARTemis-1 has been rationally designed to increase antitumor efficacy, overcome sBCMA inhibition, and incorporate the expression of a safety-gene. The generation of CARTemis-1 was successfully validated under GMP standards. A phase I/II clinical trial for patients with multiple myeloma will be conducted (EuCT number 2022-503063-15-00).

Selective inhibition of HDAC6 regulates expression of the oncogenic driver EWSR1-FLI1 through the EWSR1 promoter in Ewing sarcoma.

Ewing sarcoma (EWS) is an aggressive bone and soft tissue tumor of children and young adults in which the principal driver is a fusion gene, EWSR1-FLI1. Although the essential role of EWSR1-FLI1 protein in the regulation of oncogenesis, survival, and tumor progression processes has been described in-depth, little is known about the regulation of chimeric fusion-gene expression. Here, we demonstrate that the active nuclear HDAC6 in EWS modulates the acetylation status of specificity protein 1 (SP1), consequently regulating the SP1/P300 activator complex binding to EWSR1 and EWSR1-FLI1 promoters. Selective inhibition of HDAC6 impairs binding of the activator complex SP1/P300, thereby inducing EWSR1-FLI1 downregulation and significantly reducing its oncogenic functions. In addition, sensitivity of EWS cell lines to HDAC6 inhibition is higher than other tumor or non-tumor cell lines. High expression of HDAC6 in primary EWS tumor samples from patients correlates with a poor prognosis in two independent series accounting 279 patients. Notably, a combination treatment of a selective HDAC6 and doxorubicin (a DNA damage agent used as a standard therapy of EWS patients) dramatically inhibits tumor growth in two EWS murine xenograft models. These results could lead to suitable and promising therapeutic alternatives for patients with EWS.

Effectiveness of an Acute Care for Elders unit at a long-term care facility for frail older patients with COVID-19.

The Acute Care for Elders (ACE) is a model of care addressed to reduce the incidence of loss of self-care abilities of older adults occurring during hospitalization for acute illness. This observational study aimed to describe the effectiveness of an ACE unit at a long-term care facility to prevent functional decline (decrease in the Barthel Index score of >5 points from admission to discharge) in older adults with frailty (Clinical Frailty Scale score ≥5) and symptomatic COVID-19. Fifty-one patients (mean age: 80.2 + 9.1 years) were included. Twenty-eight (54.9%) were women, with a median Barthel index of 50 (IQR:30-60) and Charlson of 6(IQR: 5-7), and 33 (64.7%) had cognitive impairment. At discharge, 36(70.6%) patients had no functional decline, 6 (11.7%) were transferred to hospital and 4(7.8%) died. An ACE unit at a long-term care facility constitutes an alternative to hospital care to prevent hospital-associated disability for frail older patients with COVID-19.

Use of Brassica juncea and Dactylis glomerata for the phytostabilization of mine soils amended with compost or biochar.

Phytostabilization of mine soils contaminated by potentially toxic elements (PTEs) requires plants tolerant to PTE toxicity and to the poor soil physico-chemical characteristics of these areas. A pot experiment was carried out to assess the phytostabilization potential of Brassica juncea and Dactylis glomerata in mine soils amended with compost and biochar. Furthermore, the Environmental Risk of the soils and the effects of the phytostabilization process on the microbiological population size and activity in the soils were also determined. According to the Ecological Risk Index (ERI) the soils studied presented "very high risk" and As, Cd and Pb were the target elements for phytostabilization. Both amendments improved soil conditions (e.g., increasing total-N and total organic-C concentrations) and contributed to PTE (Cd, Pb and Zn) immobilization in the soil. Compost showed a more marked effect on soil microbial biomass and nutrients release in soil, which led to higher B. juncea and D. glomerata biomass in compost treated soils. Biochar treatment showed a positive effect only on D. glomerata growth, despite it provoked strong PTE immobilization in both soils. The addition of both amendments resulted in an overall reduction of PTE concentration in the plants compared to the control treatment. In addition, both plant species showed higher accumulation of PTE in the roots than in the shoots (transfer factor<1) independently of the treatment received. Therefore, they can be considered as good candidates for the phytostabilization of PTE contaminated mine soils in combination with organic amendments like biochar and compost.

The combination of epigenetic drugs SAHA and HCI-2509 synergistically inhibits EWS-FLI1 and tumor growth in Ewing sarcoma.

PURPOSE: Epigenetic regulation is crucial in mammalian development and maintenance of tissue-cell specific functions. Perturbation of epigenetic balance may lead to alterations in gene expression, resulting in cellular transformation and malignancy. Previous studies in Ewing sarcoma (ES) have shown that the Nucleosome Remodeling Deacetylase (NuRD) complex binds directly to EWS-FLI1 oncoprotein and modulates its transcriptional activity. The role of EWS-FLI1 as a driver of proliferation and transformation in ES is widely known, but the effect of epigenetic drugs on fusion activity remains poorly described. The present study evaluated the combination effects of the histone deacetylases inhibitor suberoylanilide hydroxamic acid (SAHA) and Lysine-specific demethylase1 inhibitor (HCI-2509) on different biological functions in ES and in comparison to monotherapy treatments. RESULTS: The study of proliferation and cell viability showed a synergistic effect in most ES cell lines analyzed. An enhanced effect was also observed in the induction of apoptosis, together with accumulation of cells in G1 phase and a blockage of the migratory capacity of ES cell lines. Treatment, either in monotherapy or in combination, caused a significant decrease of EWS-FLI1 mRNA and protein levels and this effect is mediated in part by fusion gene promoter regulation. The anti-tumor effect of this combination was confirmed in patient-derived xenograft mouse models, in which only the combination treatment led to a statistically significant decrease in tumor volume. CONCLUSIONS: The combination of SAHA and HCI-2509 is proposed as a novel treatment strategy for ES patients to inhibit the essential driver of this sarcoma and tumor growth.

Integrated Care on Leaving Hospital Against Medical Advice Among HIV-Infected People with Substance Use Disorders.

HIV-infected people with substance use disorders (HIV-SUDs) are at increased risk of leaving hospital against medical advice (LHAMA). The aim of this study was to evaluate the incidence of LHAMA in HIV-SUDs admitted to a patient-centered hospital where they receive integrated care, including healthcare, substance use treatment, and social support. Observational study was conducted at an urban acute-care university teaching hospital. Integrated care included a specialist in addiction medicine and a social worker incorporated into the medical staff. LHAMA was defined as participants leaving the hospital without the physician's permission and not returning within 6 h. Two hundred and ninety-nine HIV-SUDs were hospitalized, and 79 (26.4%) patients were readmitted, generating a total of 517 admissions during 2010-2016. Over the study period, 45 LHAMA were registered, yielding an incidence of 8.7%. On multiple logistic regression analysis, admission for malignancies (OR:4.2; p .02), retention in substance use treatment (OR:0.3; p .01), intravenous substance use (OR:3.1; 0.05), and marginally being foreign (OR:2.1; p .06) were independent factors associated with LHAMA. Despite the patient-centered hospital care, including integrated care, patients with lack of SUD treatment or with intravenous substance use are at increased risk of LHAMA. So, additional measures are necessary to reduce the risk of LHAMA among HIV-SUD.

[Prognostic value of assessment tools on elderly patients with chronic advanced disease and end of life, admitted to an intermediate care centre]. / Valor pronóstico de los instrumentos de evaluación de enfermedad avanzada y de final de vida en ancianos ingresados en un centro sociosanitario.

OBJECTIVE: To apply 3advanced chronic disease evaluation tools in elderly patients admitted to an intermediate and long-term care centre, and evaluate its relationship with mortality. METHODS: The NECPAL tool, PROFUND prognostic index, and Charlson comorbidity index were applied to 87 patients. RESULTS: The NECPAL tool identified 31 patients (35.6%) in need of palliative care, and according to the PROFUND index, 45 (54.7%) had high/very high risk of mortality (≥7 points), and according to Charlson index, 31 (35.6%) had high comorbidity (≥4 points). Of the NECPAL positive patients, 80.5% had a PROFUND index score ≥7, and 48.3% a Charlson index ≥ 4. These percentages were 34.4% and 28.5% in negative NECPAL patients (P<.001 and P≤.06, respectively). Correlations between the 3tools: quantitative (Spearman) number of responses in NECPAL with PROFUND (r=.57; P<.001); with Charlson (r=.214; P<.047) and between PROFUND and Charlson (r=.157; P=.148). Qualitative (kappa) NECPAL (positive/negative) with PROFUND (cut-off 6/7) (0.40; P<.001), and Charlson (cut-off 3/4) (0.19; P=.080) and between PROFUND and Charlson (0.08; P=.399). Mortality prediction (area under the curve): NECPAL 3 months 0.81 (95% CI: 0.62-1.00); 6 months 0.71 (95% CI: 0.53-0.89) and 12 months 0.67 (95% CI: 0.52-0.82). PROFUND 3 months 0.71 (95% CI: 0.50-0.91); 6 months 0.73 (95% CI: 0.58-0.87), and 12 months 0.69 (95% CI: 0.57-0.81). Charlson 3 months 0.72 (95% CI: 0.52-0.91); 6 months 0.62 (95% CI: 0.45-0.80), and 12 months 0.64 (95% CI: 0.50-0.78). CONCLUSIONS: The 3tools were significantly associated with high mortality. A low concordance was found between the results of the different tools.

Metabolic syndrome among individuals with heroin use disorders on methadone therapy: Prevalence, characteristics, and related factors.

BACKGROUND: Observational studies have reported a high prevalence of obesity and diabetes in subjects on methadone therapy; there are, however, limited data about metabolic syndrome. The aim of the study was to evaluate the prevalence of metabolic syndrome and related factors in individuals with heroin use disorder on methadone therapy. METHODS: A cross-sectional study in individuals with heroin use disorder on methadone therapy at a drug abuse outpatient center. Medical examinations and laboratory analyses after a 12-hour overnight fast were recorded. Metabolic syndrome was diagnosed according to the National Cholesterol Education Program Adult Treatment Panel III (ATP III) criteria. RESULTS: One hundred and twenty-two subjects were included, with a mean age of 46.1 ± 9 years, a median body mass index (BMI) of 25.3 kg/m2 (interquartile range [IQR]: 21.2-28), and 77.9% were men. Median exposure to methadone therapy was 13 years (IQR: 5-20). Overweight and obesity were present in 29.5% and 17.2% of the participants, respectively. Metabolic syndrome components were low high-density lipoprotein (HDL) cholesterol (51.6%), hypertriglyceridemia (36.8%), high blood pressure (36.8%), abdominal obesity (27.0%), and raised blood glucose levels (18.0%). Abdominal obesity was more prevalent in women (52% vs. 20%, P = >0.01) and high blood pressure more prevalent in men (41.1% vs. 22.2%, P = .07). Prevalence of metabolic syndrome was 29.5% (95% confidence interval [CI]: 16.6-31.8). In the multivariate logistic regression analysis, BMI (per 1 kg/m2 increase odds ratio [OR]: 1.49, 95% CI: 1.27-1.76) and exposure time to methadone therapy (per 5 years of treatment increase OR: 1.38, 95% CI: 1.28-1.48) were associated with metabolic syndrome. CONCLUSIONS: Overweight and metabolic syndrome are prevalent findings in individuals with heroin use disorder on methadone therapy. Of specific concern is the association of methadone exposure with metabolic syndrome. Preventive measures and clinical routine screening should be recommended to prevent metabolic syndrome in subjects on methadone therapy.

[Delirium prevention and treatment in elderly hip fracture]. / Prevención y tratamiento del delírium en ancianos con fractura de cadera.

The fracture of the proximal femur or hip fracture in the elderly usually happens after a fall and carries a high morbidity and mortality. One of the most common complications during hospitalization for hip fracture is the onset of delirium or acute confusional state that in elderly patients has a negative impact on the hospital stay, and prognosis, worsening functional ability, cognitive status and mortality. Also the development of delirium during hospitalization increases health care costs. Strategies to prevent and treat delirium during hospitalization for hip fracture have been less studied. In this context, this paper aims to conduct a review of the literature on strategies that exist in the prevention and treatment of delirium in elderly patients with hip fracture.

[Pharmacological treatment of dementia: when, how and for how long. Recommendations of the Working Group on Dementia of the Catalan Society of Geriatrics and Gerontology]. / Tratamiento farmacológico de la demencia: cuándo, cómo y hasta cuándo. Recomendaciones del Grupo de Trabajo de Demencias de la Sociedad Catalana de Geriatría y Gerontología.

Dementia in general--and Alzheimer's disease (AD) in particular--are bound to loom large among the most acute healthcare, social, and public health problems of the 21st century. AD shows a degenerative progression that can be slowed down--yet not halted--by today's most widely accepted specific treatments (those based on cholinesterase inhibitors as well as those using memantine). There is enough evidence to consider these treatments advisable for the mild, moderate and severe phases of the illness. However, in the final stage of the disease, a decision has to be made on whether to withdraw such treatment or not. In this paper, the Working Group on Dementia for the Catalan Society of Geriatrics and Gerontology reviews the use of these specific pharmacological treatments for AD, and, drawing on the scientific evidence thus gathered, makes a series of recommendations on when, how, and for how long, the currently existing specific pharmacological treatments should be used.

[Informing of the diagnosis in dementia]. / La información del diagnóstico en la demencia.

Dementia is a syndrome characterized by a progressive deterioration of cognitive functions, accompanied by psychiatric symptoms and behavioral disturbances that produce a progressive and irreversible disability. The way it should communicate the diagnosis of dementia is a key discussion point on which there is no unanimous agreement so far. The communicating of the diagnosis of dementia is a complex issue that affects not only, the patient but also to caregivers and health professionals who care and must conform to the ethical principles governing medical practice (autonomy, nonmaleficence, beneficence, and justice). Therefore, from the Dementia Working Group of the Catalan Geriatric Society (Grupo de Trabajo de Demencia de la Sociedad Catalana de Geriatría) arises the need to review the issue and propose a course of action for the disclosure of diagnosis.

Lower Barthel Index scores predict less prescription of pharmacological therapy in elderly patients with Alzheimer disease.

AIMS: To determine the factors associated with receiving specific treatment (cholinesterase inhibitors or/and memantine) for Alzheimer disease (AD) in elderly patients. METHODS: An observational study carried out in 289 consecutive outpatients aged >64 years with dementia. We collected data on specific AD therapy, sociodemographic variables, Barthel Index (BI), Lawton and Brody Index (LI), Mini Mental State Examination, Global Deterioration Scale (GDS), Charlson Index and the total number of drugs chronically prescribed. Patients receiving specific therapy for dementia were compared with the rest. RESULTS: Two hundred and thirty-three (80.6%) patients were receiving specific treatment for dementia, with 197 (84.5%) receiving monotherapy and the rest (15.4%) combined therapy. The bivariate analysis showed that age, marital status, place of residence, BI and LI, cognitive status and disease severity (GDS) were factors associated with receiving specific dementia therapy. Multiple stepwise logistic regression analysis showed that a lower BI (beta = -0.25; odds ratio 0.976, 95% confidence interval = 0.966-0.986; p < 0.0001) was the only factor independently associated with not receiving specific therapy for AD. CONCLUSIONS: Of the possible factors related to elderly patients receiving specific therapy for AD, a poor BI score was the most important factor associated with not receiving treatment.

[High prevalence of neuroleptic drug use in elderly people with dementia]. / Alta prevalencia de consumo de neurolépticos en las personas ancianas con demencia.

BACKGROUND AND OBJECTIVES: The management of psychological and behavioural symptoms associated with dementia frequently requires the use of neuroleptic drugs. The objective of this study was to determine the prevalence, characteristics and possible differential factors of people aged > or = 65 years with dementia who take or not neuroleptic drugs. The subgroup with Alzheimer disease was analysed. PATIENTS AND METHODS: Five-hundred and fifteen patients aged > or = 65 years with dementia were prospectively evaluated. Data were collected on sociodemographic variables, type of dementia, Barthel Index (BI), Lawton Index (LI), Mini Mental State Exam (MMSE), Charlson Index, treatment with neuroleptic, antidepressants, benzodiazepines and non-benzodiazepine hypnotic-sedatives drugs, specific dementia treatments, vascular risk factors and comorbidities. The stage and severity of dementia were evaluated by the Global Deterioration Scale (GDS), creating two groups: Mild-moderate (GDS 3, 4 and 5) and severe (GDS 6 and 7) disease. RESULTS: There were 364 women (70%) and 151 men, with a mean age of 81+/-6 years, of whom 10.1% were institutionalized. Two hundred and seventy patients (52.5%) had mild-moderate disease and 245 had severe disease (47.5%). Neuroleptic drugs were being taken by 233 (45.2%) patients. In the multivariate analysis, neuroleptic drug use was associated with male gender, institutionalization, worse LI scores, more severe dementia and not having heart failure. The subgroup with Alzheimer disease was associated with worse IB and not having arterial hypertension. CONCLUSION: A high percentage of elderly patients with dementia are treated with neuroleptic drugs. There are significant differences in the prescription of neuroleptic drugs according to patient sociodemographic characteristics, severity of dementia and comorbidities.

Neurokinin-1 receptors located in human retinoblastoma cell lines: antitumor action of its antagonist, L-732,138.

PURPOSE: The authors have recently demonstrated that substance P and L-733,060 induce cell proliferation and cell inhibition, respectively, in human retinoblastoma cell lines. However, the presence of neurokinin-1 receptors has not been demonstrated in such cell lines, nor is it known whether other neurokinin-1 receptor antagonists exert antitumoral action against retinoblastoma cell lines. The purpose of this study was to demonstrate the presence of neurokinin-1 receptors in the human retinoblastoma cell lines WERI-Rb-1 and Y-79 and to study the growth inhibitory capacity of the neurokinin-1 receptor antagonist L-732,138 against those cell lines. The authors also sought to demonstrate that the administration of L-732,138 or L-733,060 induces apoptosis in retinoblastoma cells and that neurokinin-1 receptors and substance P are present in primary retinoblastoma. METHODS: Immunoblot analysis was used to determine neurokinin-1 receptors, and a Coulter counter was used to determine viable cell numbers; this was followed by application of the tetrazolium compound WST-8, a colorimetric method, to evaluate cell viability. DAPI stain was applied to assess chromatin condensation, characteristic of apoptosis, and immunoperoxidase was used to demonstrate neurokinin-1 receptors and substance P in eyes with primary retinoblastoma. RESULTS: Neurokinin-1 receptors were present in both retinoblastoma cell lines studied. Three identical bands (isoforms of approximately 33, 58, and 75 kDa) were observed in both cell lines. Moreover, L-732,138 inhibited the growth of both cell lines studied, with and without previous administration of substance P. This inhibition occurred in a dose-dependent manner, with the IC50 values of 60.47 microM for WERI-Rb1 and 56.78 microM for Y-79. Moreover, apoptosis was observed in both cell lines after the administration of L-732,138 or L-733,060. In fixed eyes with primary retinoblastoma, a high density of neurokinin-1 receptors was observed in tumor cells, whereas a very low number of such cells contained substance P. CONCLUSIONS: This study showed that the same isoforms of the neurokinin-1 receptor are present in human retinoblastoma cell lines WERI-Rb-1 and Y-79. Both L-732,138 and L-733,060 can induce apoptosis in these cell lines and therefore can act as antitumoral agents. Primary retinoblastoma specimens display neurokinin-1 receptor immunolabeling. These results suggest that the neurokinin-1 receptor may be a promising new target for the treatment of retinoblastoma.