CAR-T cell manufacturing landscape—Lessons from the past decade and considerations for early clinical development

CAR-T cell therapies have consolidated their position over the last decade as an effective alternative to conventional chemotherapies for the treatment of a number of hematological malignancies. With an exponential increase in the number of commercial therapies and hundreds of phase 1 trials exploring CAR-T cell efficacy in different settings (including autoimmunity and solid tumors), demand for manufacturing capabilities in recent years has considerably increased. In this review, we explore the current landscape of CAR-T cell manufacturing and discuss some of the challenges limiting production capacity worldwide. We describe the latest technical developments in GMP production platform design to facilitate the delivery of a range of increasingly complex CAR-T cell products, and the challenges associated with translation of new scientific developments into clinical products for patients. We explore all aspects of the manufacturing process, namely early development, manufacturing technology, quality control, and the requirements for industrial scaling. Finally, we discuss the challenges faced as a small academic team, responsible for the delivery of a high number of innovative products to patients. We describe our experience in the setup of an effective bench-to-clinic pipeline, with a streamlined workflow, for implementation of a diverse portfolio of phase 1 trials

Strichartz Estimates for the Vibrating Plate Equation

We study the dispersive properties of the linear vibrating plate (LVP) equation. Splitting it into two Schr\"odinger-type equations we show its close relation with the Schr\"odinger equation. Then, the homogeneous Sobolev spaces appear to be the natural setting to show Strichartz-type estimates for the LVP equation. By showing a Kato-Ponce inequality for homogeneous Sobolev spaces we prove the well-posedness of the Cauchy problem for the LVP equation with time-dependent potentials. Finally, we exhibit the sharpness of our results. This is achieved by finding a suitable solution for the stationary homogeneous vibrating plate equation.Comment: 18 pages, 4 figures, some misprints correcte

Numerical analysis of deep-seated mass movements in the Magura Nappe; Flysch Belt of the Western Carpathians (Czech Republic)

Deep-seated slope failures are common features in the mountains of the Raca Unit, Magura Nappe of the Flysch Belt of Western Carpathians. Since they represent very complicated system, understanding of their evolution and triggers still remains unclear. We tried to provide a back-analysis of their development by using a finite difference code (FDM) of continua (Flac 4.0). We confirmed that such large mass movements could be triggered by water saturation of the bedrock in the three particular geological and geomorphic settings. Such situation could have been caused by heavy rainfalls in humid phases of the Holocene or permafrost melting in Late Glacial. The effects of faulting, very deep weathering of the bedrock, low geotechnical parameters of smectite-rich material and the local slope geometry have also been accounted for in numerical models, as well as the other triggering factors of slope instability. FDM modelled shear zones are in agreement with observations

The Syntaxin-1A gene single nucleotide polymorphism rs4717806 associates with the risk of ischemic heart disease

Ischemic heart disease (IHD) has a genetic predisposition and a number of cardiovascular risk factors are known to be affected by genetic factors. Development of metabolic syndrome and insulin resistance, strongly influenced by lifestyle and environmental factors, frequently occur in subjects with a genetic susceptibility. The definition of genetic factors influencing disease susceptibility would allow to identify individuals at higher risk and thus needing to be closely monitored.To this end, we focused on a complex of soluble-N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs), playing an important role in metabolic syndrome and insulin resistance, involved in endothelial dysfunction and heart disease. We assessed if genetic variants of the SNARE genes are associated with IHD.SNAP25 rs363050, Stx-1A rs4717806, rs2293489, and VAMP2 26bp ins/del genetic polymorphisms were analyzed in a cohort of 100 participants who underwent heart surgery; 56 of them were affected by IHD, while 44 were not. A statistical association of plasma glycemia and insulin resistance, calculated as Triglyceride glucose (TyG) index, was observed in IHD (P<.001 and P=.03, respectively) after binomial logistic stepwise regression analysis, adjusted by age, gender, diabetes positivity, waist circumference, and cholesterol plasma level. Among genetic polymorphisms, rs4717806(A) and rs2293489(T), as well as the rs4717806 - rs2293489 (A-T) haplotype were associated with higher risk for IHD (Pc=.02; Pc=.02; P=.04, respectively). Finally, a statistical association of rs4717806(AA) genotype with higher TyG index in IHD patients (P=.03) was highlighted by multiple regression analysis considering log-transformed biochemical parameters as dependent variable and presence of coronary artery disease, age, gender, waist circumference, presence of diabetes as predictors. These results point to a role of the Stx-1A rs4717806 SNP in IHD, possibly due to its influence on Stx-1A expression and, as a consequence, on insulin secretion and glucose metabolism

Number of implants placed for complete‐arch fixed prostheses: A systematic review and meta‐analysis

Objectives The main purpose of this systematic review was to evaluate outcomes related to the number of implants utilized to support complete‐arch fixed prostheses, both for the maxilla and the mandible. Materials and methods This review followed the reporting guidelines of the Preferred Reporting Items for Systematic Reviews and Meta‐Analyses (PRISMA). A focused question using the PICO format was developed, questioning whether “In patients with an implant supported fixed complete dental prosthesis, do implant and prosthetic survival outcomes differ between five or more compared to fewer than five supporting implants?”. A comprehensive search of the literature was formulated and performed electronically and by hand search. Two independent reviewers selected the papers and tabulated results. Primary outcomes analyzed were implant and prosthesis survival. Implant distribution, loading, and type of retention were observed as secondary outcomes, as they relate to the number of implants. A meta‐analysis was performed to compare results for studies by number of implants. Results The search strategy identified 1,579 abstracts for initial review. Based on evaluation of the abstracts, 359 articles were identified for full‐text evaluation. From these, 93 were selected and included in this review, being nine RCTs, 42 prospective and 42 retrospective. Of the 93 selected studies, 28 reported number of implants for the maxilla, 46 for the mandible, and 19 for both maxilla and mandible. The most reported number of implants for the “fewer than five” group is 4 for the maxilla, and 3 and 4 for the mandible, whereas for the “five or more” implants group, the most reported number of implants was 6 for the maxilla and 5 for the mandible. No significant differences in the primary outcomes analyzed were identified when fewer than five implants per arch were compared with five or more implants per arch (p > 0.05), in a follow‐up time ranging from 1 to 15 years (median of 8 years). Conclusions Evidence from this systematic review and meta‐analysis suggests that the use of fewer than five implants per arch, when compared to five or more implants per arch, to support a fixed prosthesis of the completely edentulous maxilla or mandible, present similar survival rates, with no statistical significant difference at a p < 0.05 and a confidence interval of 95%

Tunable control of CAR T cell activity through tetracycline mediated disruption of protein-protein interaction

Chimeric antigen receptor (CAR) T cells are a promising form of cancer immunotherapy, although they are often associated with severe toxicities. Here, we present a split-CAR design incorporating separate antigen recognition and intracellular signaling domains. These exploit the binding between the tetracycline repressor protein and a small peptide sequence (TIP) to spontaneously assemble as a functional CAR. Addition of the FDA-approved, small molecule antibiotic minocycline, acts as an "off-switch" by displacing the signaling domain and down-tuning CAR T activity. Here we describe the optimization of this split-CAR approach to generate a CAR in which cytotoxicity, cytokine secretion and proliferation can be inhibited in a dose-dependent and reversible manner. Inhibition is effective during on-going CAR T cell activation and inhibits activation and tumor control in vivo. This work shows how optimization of split-CAR structure affects function and adds a novel design allowing easy CAR inhibition through an FDA-approved small molecule

Potential of Magnetic Hyperthermia to Stimulate Localized Immune Activation

Magnetic hyperthermia (MH) harnesses the heat-releasing properties of superparamagnetic iron oxide nanoparticles (SPIONs) and has potential to stimulate immune activation in the tumor microenvironment whilst sparing surrounding normal tissues. To assess feasibility of localized MH in vivo, SPIONs are injected intratumorally and their fate tracked by Zirconium-89-positron emission tomography, histological analysis, and electron microscopy. Experiments show that an average of 49% (21-87%, n = 9) of SPIONs are retained within the tumor or immediately surrounding tissue. In situ heating is subsequently generated by exposure to an externally applied alternating magnetic field and monitored by thermal imaging. Tissue response to hyperthermia, measured by immunohistochemical image analysis, reveals specific and localized heat-shock protein expression following treatment. Tumor growth inhibition is also observed. To evaluate the potential effects of MH on the immune landscape, flow cytometry is used to characterize immune cells from excised tumors and draining lymph nodes. Results show an influx of activated cytotoxic T cells, alongside an increase in proliferating regulatory T cells, following treatment. Complementary changes are found in draining lymph nodes. In conclusion, results indicate that biologically reactive MH is achievable in vivo and can generate localized changes consistent with an anti-tumor immune response