Combination of lentiviral and genome editing technologies for the treatment of sickle cell disease.

Sickle cell disease (SCD) is caused by a mutation in the ß-globin gene leading to polymerization of the sickle hemoglobin (HbS) and deformation of red blood cells. Autologous transplantation of hematopoietic stem/progenitor cells (HSPCs) genetically modified using lentiviral vectors (LVs) to express an anti-sickling ß-globin leads to some clinical benefit in SCD patients, but it requires high-level transgene expression (i.e., high vector copy number [VCN]) to counteract HbS polymerization. Here, we developed therapeutic approaches combining LV-based gene addition and CRISPR-Cas9 strategies aimed to either knock down the sickle ß-globin and increase the incorporation of an anti-sickling globin (AS3) in hemoglobin tetramers, or to induce the expression of anti-sickling fetal γ-globins. HSPCs from SCD patients were transduced with LVs expressing AS3 and a guide RNA either targeting the endogenous ß-globin gene or regions involved in fetal hemoglobin silencing. Transfection of transduced cells with Cas9 protein resulted in high editing efficiency, elevated levels of anti-sickling hemoglobins, and rescue of the SCD phenotype at a significantly lower VCN compared to the conventional LV-based approach. This versatile platform can improve the efficacy of current gene addition approaches by combining different therapeutic strategies, thus reducing the vector amount required to achieve a therapeutic VCN and the associated genotoxicity risk.

Toward a comparison and an optimization of CT protocols using new metrics of dose and image quality part I: prediction of human observers using a model observer for detection and discrimination tasks in low-dose CT images in various scanning conditions.

In the context of reducing the patient dose coming from CT scanner examinations without penalizing the diagnosis, the assessment of both patient dose and image quality (IQ) with relevant metrics is crucial. The present study represents the first stage in a larger work, aiming to compare and optimize CT protocols using dose and IQ new metrics. We proposed here to evaluate the capacity of the Non-PreWhitening matched filter with an eye (NPWE) model observer to be a robust and accurate estimation of IQ. We focused our work on two types of clinical tasks: a low contrast detection task and a discrimination task. We designed a torso-shaped phantom, including Plastic Water®slabs with cylindrical inserts of different diameters, sections and compositions. We led a human observer study with 13 human observers on images acquired in multiple irradiation and reconstruction scanning conditions (voltage, pitch, slice thickness, noise level of the reconstruction algorithm, energy level in dual-energy mode and dose), to evaluate the behavior of the model observer compared to the human responses faced to changing conditions. The model observer presented the same trends as the human observers with generally better results. We rescaled the NPWE model on the human responses by scanning conditions (kVp, pitch, slice thickness) to obtain the best agreement between both observer types, estimated using the Bland-Altman method. The impact of some scanning parameters was estimated using the correct answer rate given by the rescaled NPWE model, for both tasks and each insert size. In particular, the comparison between the dual-energy mode at 74 keV and the single-energy mode at 120 kVp showed that, if the 120 kVp voltage provided better results for the smallest insert at the lower doses for both tasks, their responses were equivalent in many cases.