On-site Manufacturing Research Articles

Lessons to Learn for 3D Printing of Drug Products by Semisolid Extrusion (SSE)

Semisolid extrusion (SSE) 3D printing (3DP) technology is emerging due to its simplicity and potential for on-site manufacturing of personalized drug products with tailored functionality (dose, release profile), as well as recognizability (size, shape, color). However, even a minor change in the composition of the ink (the feedstock material) and the printing process parameters can largely influence the outcome of printing. This paper summarizes the recent SSE 3DP studies, where the important factors affecting the quality of the printed drug products are discussed. Further challenges are showcased by introducing a case study focusing on the design of oral theophylline immediate-release drug products. The identified crucial factors, such as the printing hardware and connected software, printing parameters, and composition of the ink are discussed. Especially, the rheological properties of the ink during the printing process, together with solidification, mechanical properties, and morphology studies of already printed products are deliberated to gain more understanding of the printability of drug products by SSE. This work aims to provide an overview of design aspects related to SSE-based fabrication of personalized drug products.

Challenges in Global Access to CAR-T cells: an Asian Perspective.

The use of cell therapy for clinical applications has seen a dramatic increase in recent years, primarily in oncology, especially with the use of chimeric antigen receptor (CAR) T-cell therapies. However, there are some barriers to the widespread adoption of CAR-T cell therapies globally, primarily because of the high cost of manufacturing these cells and clinical infrastructure considerations. We reviewed the different strategies adopted across Asia to implement CAR-T cell therapy and found that these included patient assistance programs, close engagement with funders, cost-effectiveness studies, on-site manufacturing of CAR-T cells, and joint ventures between local partners and foreign pharmaceutical companies. Although on-site manufacturing can reduce the cost of genetic engineering and expansion, it does not address many other hidden costs and quality considerations. Future growth in large-scale regional manufacturing, facilitated by cutting-edge science and innovation, could reduce costs through economies of scale and facilitate the eagerly needed global access.

Off the Grid: A new strategy for material-jet 3D printing with enhanced sub-droplet resolution

Drop-on-Demand additive manufacturing could offer a facile solution for scalable on-site manufacturing. With an increasing number of functional materials available for this technology, there are growing opportunities for applications, such as electronics. Here we report on a novel printing strategy, Off-the-Grid (OtG), which enables refined positioning of individual droplets and enhanced resolution compared to the traditional printing strategy. We demonstrate successful printing of structures with feature position control smaller than a single droplet size, and hence enhanced shape fidelity for intricate designs. This strategy is extended to filled patterns, enabling improved layer coverage and customisable inter-layer droplet positioning to control surface morphology. The OtG strategy is applied to produce functional designs, such as conformable circuitry and miniaturized antennae, and is transferable to different materials, from metal nanoparticle and polymeric inks on inkjet platforms, to molten metals on a MetalJet printer. These results could advance exploitation of AM in electronics, wearable electronics, medical devices, and metamaterials.

Application of Hybrid Laser Arc Welding for Construction of LNG Tanks Made of Thick Cryogenic 9% Ni Steel Plates

AbstractHybrid laser-arc welding (HLAW) was applied for butt welding of 14.5 mm thick plates of ferritic cryogenic steel X8Ni9 containing 9% Ni, which is used for manufacturing storage and transport facilities of liquefied natural gas (LNG). The weld seam formation and the achievable metallurgical and mechanical properties of the hybrid welds were investigated experimentally for two types of filler wire, an austenitic wire dissimilar to the base metal (BM) and an experimentally produced matching ferritic wire. Safe penetration and uniform distribution of the austenitic filler metal in the narrow hybrid weld could only be achieved in the upper, arc-dominated part of the weld. The pronounced heterogeneous distribution of the austenitic filler metal in the middle part and in the root area of the weld could not ensure sufficient notched impact toughness of the weld metal (WM). As a result, a decrease in the impact energy down to 17 ± 3 J was observed, which is below the acceptance level of ≥ 34 J for cryogenic applications. In contrast, the use of a matching ferritic filler wire resulted in satisfactory impact energy of the hybrid welds of up to 134 ± 52 J at the concerned cryogenic temperature of -196 °C. The obtained results contribute to an important and remarkable conversion in automated manufacturing of LNG facilities. In other words, the results will help to develop a new laser-based welding technology, where both quality and productivity are considered.The efficiency of the developed welding process has been demonstrated by manufacturing a prototype where a segment of the inner wall of large size LNG storage tank was constructed. In this concern, hybrid laser arc welding was conducted in both horizontal (2G) and vertical (3G) positions as a simulation to the actual onsite manufacturing. The prototype was fabricated twice where its quality was confirmed based on non-destructive and destructive examinations.

A Flow Sensing Device Formed Exclusively by Employing Additive Manufacturing for On-Site Fabrication Aboard a Ship.

This work concerns the design, fabrication, and testing of a novel air-flow sensor employing exclusively additive manufacturing that can be fabricated on-site, aboard a ship, or in a similarly remote area, without relying on external manufacturing facilities. The developed device's principle of operation is based on vortex shedding; its novelty focuses on employing solely additive manufacturing technology, for the manufacturing-in a single process step-of all the sensor's main elements. In more detail, the required flow-shaping housing, the appropriate piezoresistive sensing element, and the electrical interconnection pads are all constructed in a single process step, through standard Fused Deposition Modeling (FDM) 3D technology. Direct communication to the necessary readout circuitry can be easily achieved through standard soldering utilizing the integrated contact pads of the sensor. The prototype was preliminary characterized, validating its proper functionality. Key features of the proposed device are low cost, fast on-site manufacturing of the entire measuring device, robustness, and simplicity, suggesting numerous potential applications in the shipbuilding industry and other industrial sectors.

Development of a 3D-printed, patient-specific stereotactic system for bihemispheric deep brain stimulation

The aim of the project was to develop a patient-specific stereotactic system that allows simultaneous and thus time-saving treatment of both cerebral hemispheres and that contains all spatial axes and can be used as a disposable product. Furthermore, the goal was to reduce the size and weight of the stereotactic system compared to conventional systems to keep the strain on the patient, who is awake during the operation, to a minimum. In addition, the currently mandatory computed tomography should be avoided in order not to expose the patient to harmful X-ray radiation as well as to eliminate errors in the fusion of CT and MRI data.3D printing best meets the requirements in terms of size and weight: on the one hand, the use of plastic has considerable potential for weight reduction. On the other hand, the free choice of the individual components offers the possibility to optimize the size and shape of the stereotactic system and to adapt it to the individual circumstances while maintaining the same precision. The all-in-one stereotactic system was produced by means of the Multi Jet Fusion process. As a result, the components are highly precise, stable in use, lightweight and sterilizable. The number of individual components and interfaces, which in their interaction are potential sources of error, was significantly reduced. In addition, on-site manufacturing leads to faster availability of the system.Within the project, a patient-specific stereotaxy system was developed, printed, and assembled, which enables the execution of deep brain stimulation via only three bone anchors located on the skull. Pre-developed MRI markers, which can be screwed directly onto the bone anchors via the sleeves, eliminate the need for a CT scan completely. The fusion of the data, which is no longer required, suggests an improvement in target accuracy.

Towards clinical applicability and computational efficiency in automatic cranial implant design: An overview of the AutoImplant 2021 cranial implant design challenge.

Cranial implants are commonly used for surgical repair of craniectomy-induced skull defects. These implants are usually generated offline and may require days to weeks to be available. An automated implant design process combined with onsite manufacturing facilities can guarantee immediate implant availability and avoid secondary intervention. To address this need, the AutoImplant II challenge was organized in conjunction with MICCAI 2021, catering for the unmet clinical and computational requirements of automatic cranial implant design. The first edition of AutoImplant (AutoImplant I, 2020) demonstrated the general capabilities and effectiveness of data-driven approaches, including deep learning, for a skull shape completion task on synthetic defects. The second AutoImplant challenge (i.e., AutoImplant II, 2021) built upon the first by adding real clinical craniectomy cases as well as additional synthetic imaging data. The AutoImplant II challenge consisted of three tracks. Tracks 1 and 3 used skull images with synthetic defects to evaluate the ability of submitted approaches to generate implants that recreate the original skull shape. Track 3 consisted of the data from the first challenge (i.e., 100 cases for training, and 110 for evaluation), and Track 1 provided 570 training and 100 validation cases aimed at evaluating skull shape completion algorithms at diverse defect patterns. Track 2 also made progress over the first challenge by providing 11 clinically defective skulls and evaluating the submitted implant designs on these clinical cases. The submitted designs were evaluated quantitatively against imaging data from post-craniectomy as well as by an experienced neurosurgeon. Submissions to these challenge tasks made substantial progress in addressing issues such as generalizability, computational efficiency, data augmentation, and implant refinement. This paper serves as a comprehensive summary and comparison of the submissions to the AutoImplant II challenge. Codes and models are available at https://github.com/Jianningli/Autoimplant_II.

A novel bionic decision-making mechanism for digital twin-based manufacturing system

As an innovative smart manufacturing system, the virtual entities-based decision-making process is the most typical difference between the digital twin-based manufacturing system (DTMS) and other smart manufacturing systems. Therefore, the accuracy of virtual entity-driven decision-making is the key to affecting the system reliability of the DTMS. Normally, the manufacturing process is often accompanied by complex state changes, which are collected by the perception module of the DTMS in the form of high-dimensional information. Then, the decision-making model needs to respond to these state changes in real-time and give reasonable decision results back to physical space, which has become an important scientific issue of DTMS. To fill this gap, a novel bionic decision-making mechanism for DTMS is put forward by introducing the biological sequential learning mechanism into the decision-making process. Subsequently, the systematic decision-making process imitates biological instinct and learning behavior mechanisms to explore the short-term and long-term process of decision-making. The bionic decision-making mode formed by combining the above two modes provides adaptive decision-making in different scenarios. It is believed that the bionic decision-making mechanism can help to quickly and accurately give decision-making feedback to guide on-site manufacturing and ensure product quality and manufacturing efficiency.

Towards a better understanding of the economy and culture of the Late Bronze Age in the southeastern Baltic: Tarbiškės settlements

In 2020, rescue excavations due to construction of a pipeline connecting Poland and Lithuania took place at the Bronze Age sites Tarbiškės 1 and Tarbiškės 2, eastern Lithuania, both dated to 1050–900 cal BC. They revealed a rather homogeneous archaeological assemblage which fills a gap in the development of the Bronze Age culture and economy in the southeastern Baltic. Tarbiškės Ware, from a typological as well as chronological point of view, stands in an intermediate position linking Trzciniec culture pottery with Žalioji and Early Striated Wares. Macrobotanical analysis of charred plant remains revealed that Bronze Age people at Tarbiškės cultivated Panicum miliaceum, Hordeum vulgare and Triticum sp. The Tarbiškės sites demonstrate that early farmers used to settle areas at higher elevations with sandy soils, further from large bodies of water. They used flint and other stone tools widely and lacked bronze. Tarbiškės is the first and only ancient settlement discovered in Lithuania with a workshop for on-site manufacturing of polished stone axes with drilled holes.

On-Site Manufacturing Method for Pre-Tension U-Type Pre-Stressed Concrete Girders and Analytical Performance Verification of Anchoring Blocks Used for Applying Tension Force

Development of U-type pre-stressed girders has been attempted to increase the length of I-type girders in South Korea. However, a length of 30 m or less is common because the self-weight, according to the post-tension method, is large. In this study, the pre-tension method was applied without limiting the post-tension method to induce a reduction in self-weight and in the materials used because of the decrease in the cross section. In addition, the authors proposed an application of an on-site pre-tensioning method using the internal reaction arm of a U-type girder. A pre-stressed concrete U-type girder bridge is composed of a concrete deck slab and a composite section. Structural performance characteristics, such as resistance and rigidity, were improved compared to those of the PSC I-type girders. Construction safety is also improved in the manufacturing and installation stages, and the elongation ratio is reduced because of the reduction in the weight of the girders. Therefore, it is possible to ensure the aesthetic landscape and economic efficiency of bridges. As a result, it is expected that efficient construction will be possible with high-quality factory-made and cast-in-place members. In this study, the pre-tension method is introduced in the field, and the analytical performance of the anchoring block used for tension is verified.

Valuable recycling of LCD/OLED cullet/waste glass

Common soda lime silicate waste glass generated from the bottle manufacturing, building, and automobile industries contributes to saving of production cost and CO<sub>2</sub> emission in glass industry. There is another type of waste glass recently emerged, namely, display waste glass derived from LCD and OLED industry. There are three main sources where the waste glass is generated: 1) LCD/OLED glass manufacturers, 2) LCD/OLED panel manufacturers and 3) end-of-life LCD/OELD devices. Both first and second sources are the onsite manufacturing of LCD/OLED glass or panels in four Asian countries. Whereas, the third one is anywhere LCD/OLED devices are used. Therefore, the disposal of the end LCD/OLED waste glass from the third source is not a local but a global issue. In the present paper, the recycling possibility of such waste glass is reviewed based on literatures and their results are discussed from the viewpoint of valuable recycling.

Curing and self-healing of enzymatic construction materials using nanoparticles

Preparation of construction materials on site may offer advantages in transportation and storage. However, a major limitation of on-site preparation of some materials is the need for heat and oven desiccation for adequate curing. Here, we describe a method that allows rapid curing under ambient conditions through the addition of 0.1% iron oxide nanoparticles to a carbon-negative enzymatic construction material (ECM) to make ECM-n. Specifically, we show that a low-power laser (3W at 808 nm) can cure ECM-n to an optimal mechanical strength in 12 h, which can be compared to the 14-day period needed for in situ air drying. In addition, the incorporation of nanoparticles allows rapid self-healing of large-scale flaws and that incandescent light can be used if lasers are not available. This method establishes an on-site manufacturing capability for ECM-n and other construction materials and supports thermal controllability of the local structure in low-temperature regions. • Introduction of curing method that allows for on-site fabrication • Nanoparticles reduce the curing period in a recently developed ECM-n • Curing can be carried out using a low-power laser or other light sources • ECM-n can be used as a multifunctional thermal element Wang et al. report the application of nanoparticle methodology where particles are added to enzymatic construction material (ECM) to allow curing at ambient temperatures without the need for heat/desiccation for on-site fabrication. In addition, the material can be rapidly repaired when built-in flaws are introduced.

Improving relative density and mechanical strength of lunar regolith structures via DLP-stereolithography integrated with powder surface modification process

Three-dimensional (3D) printing is one of the most promising techniques for on-site manufacturing and is expected to become a key technology for the construction and maintenance of lunar bases. In this study, powder surface modification was integrated with digital light process (DLP)-based 3D printing to improve the mechanical properties of 3D printed lunar regolith simulant (CUG-1A) structures. A silane coupling agent (KH570) was utilised to modify the CUG-1A powder, and the underlying surface modification mechanism was elucidated. In addition, the effects of the silane coupling agent on the rheological properties, wettability, cure depth and dispersity of the slurries were investigated. It was revealed that the introduction of the silane coupling agent improved the wettability between the CUG-1A powder and photocurable resin, which effectively decreased the slurry viscosity. In addition, powder surface modification enhanced the homogeneity of CUG-1A particle distribution, which is beneficial for obtaining a uniformly distributed liquid phase during the sintering process, thus promoting the sintering densification of the printed sample. The relative density and flexural strength of the CUG-1A sample prepared by the modified powder sintered at 1100 °C reached 86.47% and 91.13±5.50 MPa, being 6.5% and 22.5% higher than the values of sample prepared by the unmodified powder, respectively. Finally, complex shaped lunar regolith simulant structures were successfully fabricated via DLP-stereolithography integrated with powder surface modification, which indicating powder surface modification is an effective way to fabricate 3D printed lunar regolith simulant components with high density and mechanical strength.

Eustress in Space: Opportunities for Plant Stressors Beyond the Earth Ecosystem

Human space exploration cannot occur without reliable provision of nutritious and palatable food to sustain physical and mental well-being. This ultimately will depend upon efficient production of food in space, with on-site manufacturing on space stations or the future human colonies on celestial bodies. Extraterrestrial environments are by their nature foreign, and exposure to various kinds of plant stressors likely cannot be avoided. But this also offers opportunities to rethink food production as a whole. We are used to the boundaries of the Earth ecosystem such as its standard temperature range, oxygen and carbon dioxide concentrations, plus diel cycles of light, and we are unfamiliar with liberating ourselves from those boundaries. However, space research, performed both in true outer space and with mimicked space conditions on Earth, can help explore plant growth from its ‘first principles’. In this sense, this perspective paper aims to highlight fundamental opportunities for plant growth in space, with a new perspective on the subject. Conditions in space are evidently demanding for plant growth, and this produces “stress”. Yet, this stress can be seen as positive or negative. With the positive view, we discuss whether plant production systems could proactively leverage stresses instead of always combatting against them. With an engineering view, we focus, in particular, on the opportunities associated with radiation exposure (visible light, UV, gamma, cosmic). Rather than adapting Earth conditions into space, we advocate on rethinking the whole issue; we propose there are opportunities to exploit space conditions, commonly seen as threats, to benefit space farming.

Metal additive manufacturing of conformal cooling channels in plastic injection molds with high number of design variables

Metal additive manufacturing (MAM) is an effective way to fabricate conformal cooling channels (CCCs), which follow the curves of the plastic product in the mold body of the plastic injection. CCCs have free-curved pathways thanks to the design & manufacturing flexibilities of the MAM process so that they can achieve better cooling performance with shorter cooling time and smaller temperature non-uniformity. On the other hand, the flexibilities of the MAM process bring multiple options for design variables and the high number of design variables make the final design of CCCs complex, high-cost, and time-consuming. Considering this challenge, this study presents the entire process of MAM of CCCs for a target product with eight different design variables, which makes it a product with a high number of design variables, from the initial design to the on-site manufacturing including the steps of computer-aided design & simulations, metamodel, multiobjective optimization, and the printing quality monitoring. The target product has three main objectives that are i) the temperature difference between the maximum and minimum values at the internal wall of the mold, ii) maximum temperature in the mold body, and iii) pressure drop. The optimized product is then printed via direct metal laser sintering (DMLS) machine and the quality check is done via X-ray computed tomography.

Design of a Mass Air Flow Sensor Employing Additive Manufacturing and Standard Airfoil Geometry

This work concerns the design, fabrication, and preliminary characterization of a novel sensor for determining the air intake of low and medium power internal combustion engines employed at various applications in the marine industry. The novelty of the presented sensor focuses on the fabrication process, which is based on additive manufacturing combined with PCB technology, and the design of the sensing elements housing geometry, which is derived through suitable CFD simulations and is based on standard airfoil geometry. The proposed process enables low-cost, fast fabrication, effective thermal isolation, and facile electrical interconnection to the corresponding circuitry of the sensor. For initial characterization purposes, the prototype device was integrated into a DIESEL engine testbed while a commercially available mass air flow sensor was employed as a reference; the proper functionality of the developed prototype has been validated. Key features of the proposed device are low-cost, fast on-site manufacturing of the device, robustness, and simplicity, suggesting numerous potential applications in marine engineering.

Investigating Tissue Mechanics in vitro Using Untethered Soft Robotic Microdevices.

This paper presents the design, fabrication, and operation of a soft robotic compression device that is remotely powered by laser illumination. We combined the rapid and wireless response of hybrid nanomaterials with state-of-the-art microengineering techniques to develop machinery that can apply physiologically relevant mechanical loading. The passive hydrogel structures that constitute the compliant skeleton of the machines were fabricated using single-step in situ polymerization process and directly incorporated around the actuators without further assembly steps. Experimentally validated computational models guided the design of the compression mechanism. We incorporated a cantilever beam to the prototype for life-time monitoring of mechanical properties of cell clusters on optical microscopes. The mechanical and biochemical compatibility of the chosen materials with living cells together with the on-site manufacturing process enable seamless interfacing of soft robotic devices with biological specimen.

Development of immediate release 3D-printed dosage forms for a poorly water-soluble drug by fused deposition modeling: Study of morphology, solid state and dissolution

3D-printing technologies such as Fused Deposition Modeling (FDM) bring a unique opportunity for personalized and flexible near-patient production of pharmaceuticals, potentially improving safety and efficacy for some medications. However, FDM-printed tablets often exhibit tendency for slow dissolution due to polymer erosion-based dissolution mechanisms. Development of immediate release (IR) 3D-printed dosage with poorly water-soluble compounds is even more challenging but necessary to ensure wide applicability of the technology within pharmaceutical development portfolios. In this work, process and morphology were considered to achieve IR of BCS class IV compound lumefantrine as model active pharmaceutical ingredient (API) using basic butylated methacrylate copolymer (Eudragit EPO) as matrix former, as well as hydrophilic plasticizer xylitol and pore former maltodextrin. Grid-designed tablets with size acceptable for children from 6 years old and varying programmed infill density were successfully 3D-printed with 5% lumefantrine while higher drug load led to increased brittleness which is incompatible with 3D-printing. Lumefantrine assay was 92 to 97.5% of theoretical content depending on drug load and process parameters. 3D-printed tablets with 65% infill density met rapid release criteria, while 80% and 100% showed slower dissolution. Structural characteristics of 3D-printed tablets with non-continuous surface such as accessible porosity and specific surface area by weight and by volume were quantified by a non-destructive automated µCT-based methodology and were found to correlate with dissolution rate. Increase in accessible porosity, total surface area, specific surface area and decrease in relative density were statistically significant critical factors for modification of lumefantrine dissolution rate. Crystallinity in manufactured tablets and filaments was explored by highly sensitive Raman mapping technique. Lumefantrine was present in the fully amorphous state in the tablets exhibiting adequate stability for on-site manufacturing. The study demonstrates feasibility of immediate release FDM-3D-printed tablets with BCS class IV API and illustrates the correlation of FDM design parameters with morphological and dissolution characteristics of manufactured tablets.

A new paradigm in a low-rise housing construction

The article highlights the structure of consumer demand in the individual real estate market. Construction technologies based on "prefab housing" (construction of houses from prefabricated factory structures) are described. The transition from on-site manufacturing technologies (erection of houses directly at construction sites) to modern construction methods ("off-site manufacturing" house building) is described.

Single-Cell RNA Sequencing Identifies Expression Patterns Associated with Clinical Responses to Dual-Targeted CAR-T Cell Therapy

INTRODUCTION: Chimeric Antigen Receptor (CAR)-T cell therapy is emerging as a powerful treatment for relapsed or refractory B cell lymphomas. However, a variety of escape mechanisms prevent CAR-T cell therapy from being more uniformly effective. To better understand mechanisms of CAR-T failure among patients treated with dual-targeted CAR-T cells, we performed single-cell RNA sequencing of samples from a Phase 1 trial (NCT03019055). The clinical trial used anti-CD20, anti-CD19 CAR-T cells for the treatment of relapsed/refractory B-cell non-Hodgkin Lymphoma. Clinical responses from this study are reported independently (Shah et al. in press in Nat Med). While robust clinical responses occurred, not all patients had similar outcomes. In single-antigen specific CAR-T cells, mechanisms of resistance include antigen down-regulation, phenotype switch, or PD-1 inhibition (Song et al. Int J Mol Sci 2019). However, very little is understood about the mechanisms of failure that are specific to dual-targeted CAR-T cells. Interestingly, loss of CD19 antigen was not observed in treatment failures in the study. METHODS: De-identified patient samples were obtained as peripheral blood mononuclear cells on the day of harvest ("pre" samples), at the peak of in vivo CAR-T cell expansion which varied from day 10 to day 21 after infusion ("peak" samples), and on day 28 post-infusion ("d28" samples). The CAR-T cell infusion product was obtained on day 14 of on-site manufacturing ("product" samples). All samples were cryopreserved and single cell preparation was performed with batched samples using 10X Genomics kits. Subsequent analysis was performed in R studio using the Seurat package (Butler et al. Nat Biotech 2018) with SingleR being used to identify cell types in an unbiased manner (Aran et al. Nat Immunol 2019). RESULTS: We found that distinct T cell clusters were similarly represented in the responder and non-responder samples. The patients' clinical responses did not depend on the level of CAR expression or the percentage of CAR+ cells in the infusion product. At day 28, however, there was a considerable decrease in the percentage of CAR+ cells in the responder samples possibly due to contracture of the CAR+ T cell compartment after successful clearance of antigen-positive cells. In all samples, the CAR-T cell population shifted from a CD4+ to a CD8+ T cell predominant population after infusion. We performed differentially-expressed gene analyses (DEG) of the total and CAR-T cells. In the pre samples, genes associated with T-cell stimulation and cell-mediated cytotoxicity were highly expressed in the responder samples. Since the responders had an effective anti-tumor response, we expected these pathways to also be enriched for in the peak samples; however, this was not the case. We hypothesize that differential expression of the above genes was masked due to homeostatic expansion of the T cells following conditioning chemotherapy. Based on the DEG results, we next interrogated specific genes associated with cytotoxicity, T cell co-stimulation, and checkpoint protein inhibition. Cytotoxicity-associated genes were highly expressed among responder CD8+ T cells in the pre samples, but not in the other samples (Figure 1). Few differences were seen in specific co-stimulatory and checkpoint inhibitor genes at any timepoint in the T cell clusters. We performed gene set enrichment analyses (GSEA). Gene sets representing TCR, IFN-gamma, and PD-1 signaling were significantly increased in the pre samples of the responders but not at later time points or in the infusion products. DISCUSSION: We found a correlation between expression of genes associated with T cell stimulation and cytotoxicity in pre-treatment patient samples and subsequent response to CAR-T cell therapy. This demonstrates that the existing transcriptome of T cells prior to CAR transduction critically shapes anti-tumor responses. Further work will discover biomarkers that can be used to select patients expected to have better clinical outcomes. Figure 1 Disclosures Johnson: Miltenyi Biotec: Research Funding; Cell Vault: Research Funding. Schneider:Lentigen, a Miltenyi Biotec Company: Current Employment, Patents & Royalties. Dropulic:Lentigen, a Miltenyi Biotec Company: Current Employment, Patents & Royalties: CAR-T immunotherapy. Hari:BMS: Consultancy; Amgen: Consultancy; GSK: Consultancy; Janssen: Consultancy; Incyte Corporation: Consultancy; Takeda: Consultancy. Shah:Incyte: Consultancy; Cell Vault: Research Funding; Lily: Consultancy, Honoraria; Kite Pharma: Consultancy, Honoraria; Verastim: Consultancy; TG Therapeutics: Consultancy; Celgene: Consultancy, Honoraria; Miltenyi Biotec: Honoraria, Research Funding.