Erratum: Physiological lentiviral vectors for the generation of improved CAR-T cells.

[This corrects the article DOI: 10.1016/j.omto.2022.05.003.].

Physiological lentiviral vectors for the generation of improved CAR-T cells.

Anti-CD19 chimeric antigen receptor (CAR)-T cells have achieved impressive outcomes for the treatment of relapsed and refractory B-lineage neoplasms. However, important limitations still remain due to severe adverse events (i.e., cytokine release syndrome and neuroinflammation) and relapse of 40%-50% of the treated patients. Most CAR-T cells are generated using retroviral vectors with strong promoters that lead to high CAR expression levels, tonic signaling, premature exhaustion, and overstimulation, reducing efficacy and increasing side effects. Here, we show that lentiviral vectors (LVs) expressing the transgene through a WAS gene promoter (AW-LVs) closely mimic the T cell receptor (TCR)/CD3 expression kinetic upon stimulation. These AW-LVs can generate improved CAR-T cells as a consequence of their moderate and TCR-like expression profile. Compared with CAR-T cells generated with human elongation factor α (EF1α)-driven-LVs, AW-CAR-T cells exhibited lower tonic signaling, higher proportion of naive and stem cell memory T cells, less exhausted phenotype, and milder secretion of tumor necrosis factor alpha (TNF-α) and interferon (IFN)-É£ after efficient destruction of CD19+ lymphoma cells, both in vitro and in vivo. Moreover, we also showed their improved efficiency using an in vitro CD19+ pancreatic tumor model. We finally demonstrated the feasibility of large-scale manufacturing of AW-CAR-T cells in guanosine monophosphate (GMP)-like conditions. Based on these data, we propose the use of AW-LVs for the generation of improved CAR-T products.

Using Gene Editing Approaches to Fine-Tune the Immune System.

Genome editing technologies not only provide unprecedented opportunities to study basic cellular system functionality but also improve the outcomes of several clinical applications. In this review, we analyze various gene editing techniques used to fine-tune immune systems from a basic research and clinical perspective. We discuss recent advances in the development of programmable nucleases, such as zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeat (CRISPR)-Cas-associated nucleases. We also discuss the use of programmable nucleases and their derivative reagents such as base editing tools to engineer immune cells via gene disruption, insertion, and rewriting of T cells and other immune components, such natural killers (NKs) and hematopoietic stem and progenitor cells (HSPCs). In addition, with regard to chimeric antigen receptors (CARs), we describe how different gene editing tools enable healthy donor cells to be used in CAR T therapy instead of autologous cells without risking graft-versus-host disease or rejection, leading to reduced adoptive cell therapy costs and instant treatment availability for patients. We pay particular attention to the delivery of therapeutic transgenes, such as CARs, to endogenous loci which prevents collateral damage and increases therapeutic effectiveness. Finally, we review creative innovations, including immune system repurposing, that facilitate safe and efficient genome surgery within the framework of clinical cancer immunotherapies.

Intramyocardial bone marrow mononuclear cells versus bone marrow-derived and adipose mesenchymal cells in a rat model of dilated cardiomyopathy.

BACKGROUND: Effects of cell therapy on dilated cardiomyopathy (DCM) have been investigated in pre-clinical models using distinct cellular types in each study. A single study that compares the effectiveness of different cells is lacking. METHODS: We have compared the effects of intramyocardial injection (IMI) of bone marrow (BM)-derived mononuclear cells (MNCs), BM and adipose tissue (AT) mesenchymal stromal cells (BM-MSCs and AT-MSCs) on heart function, histological changes and myocardial ultrastructure in a rat model of DCM. Isogenic Wistar rats were used to isolate the different cell types and to induce DCM by autoimmune myocarditis. Animals were randomly assigned to receive BM-MNCs, BM-MSCs, AT-MSCs or placebo at day 42 by IMI. Serial echocardiography was used to assess cardiac function and hearts obtained after sacrifice at day 70, were used for histological and ultrastructural analysis. Serum levels of type B-natriuretic peptide (BNP) and vascular endothelial growth-factor (VEGF) were determined at different time points. RESULTS: BM-MSC treatment induced significant improvement in ejection fraction (EF), fractional shortening (FS), left ventricular systolic diameter (LVESD) and systolic volume (LVESV). In contrast, changes in echocardiographic parameters with respect to pre-treatment values in animals receiving placebo, AT-MSCs or BM-MNCs were not statistically significant. EF and FS in animals receiving AT-MSCs were superior to those receiving placebo. BM-MSC transplantation induced also improvement in cardiac fibers organization and capillary density, fibrotic tissue reduction, increase in final VEGF concentration and BNP decrease. DISCUSSION: IMI of BM or AT-MSCs improves LV function and induces more angiogenesis processes than BM-MNCs. In addition, BM-MSCs showed more anti-fibrotic effects and more ability to reorganize myocardial tissue compared with the other cell types.

Familial aortic coarctation: a rare cause of refractory hypertension in the elderly: a case report.

We report the first case of late presentation of familial aortic coarctation, a rare cause of hypertension. Diagnosis of familial aortic coarctation in the elderly is exceptional, given that in the absence of endovascular or surgical repair patients do not usually survive beyond 50 years of age. Our case concerns a 72-year-old woman with hypertension of long evolution, control of which improved markedly after endovascular repair of the coarctation. Her son had undergone surgery for repair of aortic coarctation at the age of 23 years.

Neuromodulación sacra en el tratamiento de la incontinencia fecal grave: resultados tras 10 años de experiencia / Sacral nerve stimulation for the treatment of severe faecal incontinence: Results after 10 years experience

INTRODUCCIÓN: El objetivo del estudio es presentar nuestra experiencia en el tratamiento de la incontinencia fecal grave mediante neuromodulación sacra tras los primeros 10 años de utilización de la técnica. MATERIAL Y MÉTODOS: Cuarenta y nueve pacientes con incontinencia fecal grave se trataron con neuromodulación sacra durante el periodo 2001-2011. Se practicó manometría anorrectal, ecografía endoanal y estudio de latencia pudenda. Se evaluaron diario evacuatorio, escalas de gravedad y calidad de vida de incontinencia fecal en el preoperatorio y al final del seguimiento. RESULTADOS: Un tercio de los pacientes presentó morbilidad, la mayoría leve. Se explantaron 4 dispositivos definitivos. Con una mediana de seguimiento de 37 meses, la gravedad de la incontinencia fecal, urgencia e incontinencia mejoraron significativamente al cierre del estudio. El subgrupo de pacientes con seguimiento mayor de 5 años mejoró significativamente la gravedad de la incontinencia pero no los parámetros del diario evacuatorio. No hubo diferencias significativas en la calidad de vida. Los datos descriptivos en pacientes con defectos esfinterianos no muestran peores resultados que en la integridad esfinteriana. CONCLUSIONES: La neuromodulación sacra es una técnica segura para la incontinencia fecal grave con buenos resultados funcionales a medio plazo. A largo plazo, existe también mejoría de la gravedad de la misma pero son necesarios estudios con mayor muestra para objetivar si otros parámetros clínicos y la calidad de vida avalan este dato. Los resultados preliminares en pacientes con defectos esfinterianos sugieren que la técnica puede ser eficaz en este grupo pero futuros estudios deberán confirmar estos hallazgos

INTRODUCTION: The objective of this study was to report our experience with sacral nerve stimulation for the treatment of severe faecal incontinence after the first 10 years with this technique. MATERIALS AND METHODS: Between 2001 and 2011, 49 patients with severe faecal incontinence underwent sacral nerve stimulation. Anorectal manometry, endoanal ultrasound and pudendal nerve latency were performed. Bowel habit diary, severity of faecal incontinence and quality of life scales were evaluated preoperatively and at the end of follow-up. RESULTS: Morbidity occurred in a third of patients, mostly minor. Four definitive devices were explanted. With a median follow-up of 37 months, severity of faecal incontinence, urge and incontinence episodes significantly improved at the end of follow-up. Patients' subgroup with a major follow-up of 5 years significantly improved the severity of faecal incontinence but not the parameters of the bowel habit diary. Quality of life showed no significant improvement. Descriptive data of patients with sphincter defects did not show worse results than that of patients with sphincter integrity. CONCLUSION: Sacral nerve stimulation is a safe technique for severe faecal incontinence with good functional medium-term results. In the long term, severity of the faecal incontinence also improves, but studies with larger sample are necessary to show if other clinical parameters and the quality of life support this information. Preliminary results in patients with sphincter defects suggest that this technique could be effective in this group, but future studies will have to confirm these findings

Sacral nerve stimulation for the treatment of severe faecal incontinence: results after 10 years experience.

INTRODUCTION: The objective of this study is to report our experience with sacral nerve stimulation for the treatment of severe faecal incontinence after the first 10 years with this technique. MATERIAL AND METHODS: Between 2001 and 2011, 49 patients with severe faecal incontinence underwent sacral nerve stimulation. Anorectal manometry, endoanal ultrasound and pudendal nerve latency were performed. Bowel habit diary, severity of faecal incontinence and quality of life scales were evaluated preoperatively and at the end of follow-up. RESULTS: Morbidity occurred in a third of patients, mostly minor. Four definitive devices were explanted. With a median follow-up of 37 months, severity of faecal incontinence, urge and incontinence episodes significantly improved at the end of follow-up. Patients' subgroup with major follow-up of 5 years significantly improved the severity of faecal incontinence but not the parameters of the bowel habit diary. Quality of life showed no significant improvement. Descriptive data in patients with sphincter defects did not show worse results than with sphincter integrity. CONCLUSION: Sacral nerve stimulation is a safe technique for severe faecal incontinence with good functional medium-term results. In the long term, severity of the faecal incontinence also improves but studies with larger sample are necessary to show if other clinical parameters and the quality of life support this information. Preliminary results in patients with sphincter defects suggest that this technique could be effective in this group but future studies will have to confirm these findings.

Metabolic syndrome and vascular risk estimation in a Mediterranean non-diabetic population without cardiovascular disease.

AIMS: Vascular risk equations are tools used to help prevent cardiovascular events. Our aim was to compare the REGICOR and SCORE equations in a general population and in persons with the metabolic syndrome (MS) according to the criteria of the International Diabetes Federation. METHODS AND RESULTS: We calculated the cardiovascular risk with both equations in a random sample of 838 non-diabetic persons aged 40-65years without a history of cardiovascular disease, of whom 251 had the MS. Of the 838 persons, 3.6% had a high risk according to SCORE and 1.5% according to REGICOR, and of these, 53.3% and 61.5%, respectively, had the MS. The mean risk was greater in the persons with the MS than those without (REGICOR 4.6% vs. 2.6% and SCORE 1.7% vs. 1%; p<0.01 for each). In comparison with the group without the MS, the percentage of persons with the MS who had a high risk was greater with both scales: REGICOR (3.2% vs. 0.8%, p=0.027) and SCORE (6.4% vs. 2.4%, p=0.004). The agreement (kappa index) classifying the subjects with a high risk, was 0.453 in the overall sample and 0.391 in the subgroup with the MS. CONCLUSIONS: The percentage of persons classified as having a high cardiovascular risk differed between REGICOR and SCORE. Using these scales only a small percentage of non-diabetic persons with the MS have a high risk. The presence of the MS multiplies the percentage of non-diabetic persons with a high vascular risk two-fold with SCORE and four-fold with REGICOR.

Necrosis cutánea por interferón alfa en paciente con melanoma / Cutaneous necrosis due to interferon alpha in a patient with melanoma

Presentamos el caso de un varón de 55 años con un melanoma en la región retroauricular derecha que después de un tratamiento quirúrgico y adyuvante con interferón (IFN alfa 2b) según la pauta propuesta por Kirkwood presentó, en el último mes de tratamiento, una necrosis cutánea sólo en uno de los sitios de inyección. La aparición de necrosis cutánea es una situación rara que se ha descrito en pacientes tratados con distintos tipos de IFN

We present the case of a 55-year-old male with melanoma in the right retroauricular region who had cutaneous necrosis only in one of the injection sites after adjuvant treatment with interferon (IFN alpha 2b) according to the Kirkwood proposed regime. Appearance of cutaneous necrosis is a rare condition that has been described in patients treated with different types of IFN