Preclinical efficacy and safety evaluation of interleukin-6-knockdown CAR-T cells targeting at CD19.

BACKGROUND: ssCART-19 cells with shRNA-IL-6 gene knockdown were subjected to a comprehensive safety evaluation, including efficacy, toxicity and biodistribution studies in NSG (PrkdcscidIL2rgtm1 /Bcgen) mice. METHODS: NSG mice were administered Raji-Luc and then singly dosed with ssCART-19 cells via intravenous infusion. ssCART-19 DNA fragments were quantified in different tissues by qPCR, and the optical intensity of Raji-Luc was determined for evaluate the efficacy of regular CAR-T and ssCART-19 cells. In toxicity study, clinical symptoms observation, body weight measurements, serum biochemical analysis, human cytokine detection, lymphocytes subsets quantification, necropsy and histopathological examination were performed. RESULTS: The ssCART-19 DNA was mainly concentrated in the liver within 3 hours, and was widely distributed in most of the organs/tissues for 4 weeks after administration. Chimeric antigen receptor gene modified T cells (CAR-Ts) were detected in the peripheral blood with a significant increase in number beginning at approximately 3 weeks. ssCART-19 administration resulted in increased of interferon-gamma (IFN-γ), tumor necrosis factor (TNF), interleukin-2 (IL-2), and IL-17A and decreased IL-10 and IL-6 levels. ssCART-19 inhibited the proliferation of Raji-Luc cells in tumor-bearing NSG mice, and reduced the incidence of lymphomas in the liver, kidneys and spleen. It alleviated clinical symptoms caused by tumor cell proliferation in treated animals. CONCLUSIONS: ssCART-19 prolongs the survival time of tumor-bearing mice without obvious risks of immunotoxicity and tumorigenicity. ssCART-19 DNA was found in the brains of treated animals, however no significant central nervous system toxicity was observed. These data were used to support an investigational new drug (IND) application of ssCART-19 for clinical trial in China.

Triptolide induces oxidative damage in NRK-52E cells through facilitating Nrf2 degradation by ubiquitination via the GSK-3ß/Fyn pathway.

Triptolide (TP) isolated from Tripterygium wilfordii Hook F. (TWHF) shows extensive anti-inflammation, immunosuppression and anti-tumor properties. However, its therapeutic potential is limited by its severe side effects, especially the nephrotoxicity. This study intended to explore the role of the GSK-3ß/Fyn pathway in TP-induced oxidative damage and the potential mechanism of Nrf2 protein downregulation. Our data showed that TP induced oxidative stress and cell damage in the rat renal tubular epithelial cell line NRK-52E cells by activation of GSK-3ß and nuclear translocation of Fyn, which resulted in decreased Nrf2 nuclear translocation. Moreover, TP significantly induced Nrf2 degradation by ubiquitination, which was blocked by the proteasome inhibitor MG132. In addition, cotreatment with a typical GSK-3ß inhibitor, lithium chloride, promoted the nuclear translocation of Nrf2 and decreased the nuclear translocation of Fyn, which led to reduced cell damage, LDH leakage, glutathione depletion and cell apoptosis. Collectively, our results indicated that TP induced oxidative damage in NRK-52E cells by facilitating Nrf2 degradation by ubiquitination via the GSK-3ß/Fyn pathway.

Clinical significance of optical coherence tomography-guided angioplasty on treatment selection.

The present study aimed to observe whether optical coherence tomography (OCT)-guided angioplasty is able to provide useful clinical information beyond that obtained by angiography as well as provide recommendations for physicians that may improve treatment selection. This prospective study included 83 patients with coronary artery disease (>18 years) undergoing coronary angiography (CAG) for ST-elevation myocardial infarction (n=13), non-ST-elevation myocardial infarction (n=19), stable angina (n=22), unstable angina (n=10), silent ischemia (n=11), or elective percutaneous coronary intervention (n=8). Following the initial CAG (CAG-pre), the patients underwent OCT before angioplasty (OCT-pre, 24 patients), after angioplasty (OCT-post, 22 patients), or both (37 patients). The thrombus burden, calcification and plaque dissection or rupture were compared between the OCT-pre and CAG-pre recordings. Following angioplasty, stent malapposition, suboptimal stent deployment, suboptimal stent lesion coverage, and edge dissection were compared between OCT-post and CAG-post alone. Among the 83 patients, 45.7% had single-vessel and 54.3% had multiple-vessel disease. OCT pre- and post-angioplasty revealed significantly more information on the procedure than CAG alone. This clinical information changed the clinical strategies in 41/83 (49.4%) patients, including 58 modifications of therapeutic strategy (69.9%, 58/83): Thrombus aspiration in 2 cases (2.4%), administration of glycoprotein IIb/IIIa inhibitors in 8 cases (9.6%), additional balloon inflation in 23 cases (27.7%), additional stent implantation in 17 cases (20.5%), avoiding stent implantation in 4 cases (4.8%), collateral intervention in 2 cases (2.4%), and guidewire reposition in 2 cases (2.4%). In conclusion, OCT-pre and OCT-post provided additional clinical information beyond that obtained by angiography alone, which resulted in modification of the treatment strategies in half of the included patients.