Dietary thiamine influences l-asparaginase sensitivity in a subset of leukemia cells.

Tumor environment influences anticancer therapy response but which extracellular nutrients affect drug sensitivity is largely unknown. Using functional genomics, we determine modifiers of l-asparaginase (ASNase) response and identify thiamine pyrophosphate kinase 1 as a metabolic dependency under ASNase treatment. While thiamine is generally not limiting for cell proliferation, a DNA-barcode competition assay identifies leukemia cell lines that grow suboptimally under low thiamine and are characterized by low expression of solute carrier family 19 member 2 (SLC19A2), a thiamine transporter. SLC19A2 is necessary for optimal growth and ASNase resistance, when standard medium thiamine is lowered ~100-fold to human plasma concentrations. In addition, humanizing blood thiamine content of mice through diet sensitizes SLC19A2-low leukemia cells to ASNase in vivo. Together, our work reveals that thiamine utilization is a determinant of ASNase response for some cancer cells and that oversupplying vitamins may affect therapeutic response in leukemia.

ZBTB1 Regulates Asparagine Synthesis and Leukemia Cell Response to L-Asparaginase.

Activating transcription factor 4 (ATF4) is a master transcriptional regulator of the integrated stress response (ISR) that enables cell survival under nutrient stress. The mechanisms by which ATF4 couples metabolic stresses to specific transcriptional outputs remain unknown. Using functional genomics, we identified transcription factors that regulate the responses to distinct amino acid deprivation conditions. While ATF4 is universally required under amino acid starvation, our screens yielded a transcription factor, Zinc Finger and BTB domain-containing protein 1 (ZBTB1), as uniquely essential under asparagine deprivation. ZBTB1 knockout cells are unable to synthesize asparagine due to reduced expression of asparagine synthetase (ASNS), the enzyme responsible for asparagine synthesis. Mechanistically, ZBTB1 binds to the ASNS promoter and promotes ASNS transcription. Finally, loss of ZBTB1 sensitizes therapy-resistant T cell leukemia cells to L-asparaginase, a chemotherapeutic that depletes serum asparagine. Our work reveals a critical regulator of the nutrient stress response that may be of therapeutic value.

Targeting extracellular nutrient dependencies of cancer cells.

BACKGROUND: Cancer cells rewire their metabolism to meet the energetic and biosynthetic demands of their high proliferation rates and environment. Metabolic reprogramming of cancer cells may result in strong dependencies on nutrients that could be exploited for therapy. While these dependencies may be in part due to the nutrient environment of tumors, mutations or expression changes in metabolic genes also reprogram metabolic pathways and create addictions to extracellular nutrients. SCOPE OF REVIEW: This review summarizes the major nutrient dependencies of cancer cells focusing on their discovery and potential mechanisms by which metabolites become limiting for tumor growth. We further detail available therapeutic interventions based on these metabolic features and highlight opportunities for restricting nutrient availability as an anti-cancer strategy. MAJOR CONCLUSIONS: Strategies to limit nutrients required for tumor growth using dietary interventions or nutrient degrading enzymes have previously been suggested for cancer therapy. The best clinical example of exploiting cancer nutrient dependencies is the treatment of leukemia with l-asparaginase, a first-line chemotherapeutic that depletes serum asparagine. Despite the success of nutrient starvation in blood cancers, it remains unclear whether this approach could be extended to other solid tumors. Systematic studies to identify nutrient dependencies unique to individual tumor types have the potential to discover targets for therapy.

Squalene accumulation in cholesterol auxotrophic lymphomas prevents oxidative cell death.

Cholesterol is essential for cells to grow and proliferate. Normal mammalian cells meet their need for cholesterol through its uptake or de novo synthesis1, but the extent to which cancer cells rely on each of these pathways remains poorly understood. Here, using a competitive proliferation assay on a pooled collection of DNA-barcoded cell lines, we identify a subset of cancer cells that is auxotrophic for cholesterol and thus highly dependent on its uptake. Through metabolic gene expression analysis, we pinpoint the loss of squalene monooxygenase expression as a cause of cholesterol auxotrophy, particularly in ALK+ anaplastic large cell lymphoma (ALCL) cell lines and primary tumours. Squalene monooxygenase catalyses the oxidation of squalene to 2,3-oxidosqualene in the cholesterol synthesis pathway and its loss results in accumulation of the upstream metabolite squalene, which is normally undetectable. In ALK+ ALCLs, squalene alters the cellular lipid profile and protects cancer cells from ferroptotic cell death, providing a growth advantage under conditions of oxidative stress and in tumour xenografts. Finally, a CRISPR-based genetic screen identified cholesterol uptake by the low-density lipoprotein receptor as essential for the growth of ALCL cells in culture and as patient-derived xenografts. This work reveals that the cholesterol auxotrophy of ALCLs is a targetable liability and, more broadly, that systematic approaches can be used to identify nutrient dependencies unique to individual cancer types.

Immunogenicity of pulsatile-release PLGA microspheres for single-injection vaccination.

The World Health Organization's Expanded Programme on Immunization has led to a dramatic rise in worldwide vaccination rates over the past 40years, yet 19.4 million infants remain underimmunized each year. Many of these infants have received at least one vaccine dose but may remain unprotected because they did not receive subsequent booster doses due to logistical challenges. This study aimed to develop injectable controlled release microparticles with kinetics that mimic common vaccine dosing regimens consisting of large antigen doses administered periodically over the course of months in order to eliminate the need for boosters. Sixteen poly(lactic-co-glycolic acid) (PLGA) microsphere formulations containing bovine serum albumin (BSA) as a model vaccine antigen were screened in vitro to determine their respective release kinetics. Three formulations that exhibited desirable pulsatile release profiles were then selected for studying immunogenicity in mice. Two low-dose microsphere formulations induced peak anti-BSA IgG antibody titers of 13.9±1.3 and 13.7±2.2 log2 compared to 15.5±1.5 log2 for a series of three bolus injections delivered at 0, 4, and 8weeks with an equivalent cumulative dose. Similarly, high-dose formulations induced peak antibody titers that were 16.1±2.1 log2 compared to 17.7±2.2 log2 for controls. All three microparticle formulations studied in vivo induced peak antibody titers that were statistically similar to bolus controls. These results suggest that pulsatile antigen release from polymeric microparticles is a promising approach for single-injection vaccination, which could potentially reduce the logistical burden associated with immunization in the developing world.

Fabrication of fillable microparticles and other complex 3D microstructures.

Three-dimensional (3D) microstructures created by microfabrication and additive manufacturing have demonstrated value across a number of fields, ranging from biomedicine to microelectronics. However, the techniques used to create these devices each have their own characteristic set of advantages and limitations with regards to resolution, material compatibility, and geometrical constraints that determine the types of microstructures that can be formed. We describe a microfabrication method, termed StampEd Assembly of polymer Layers (SEAL), and create injectable pulsatile drug-delivery microparticles, pH sensors, and 3D microfluidic devices that we could not produce using traditional 3D printing. SEAL allows us to generate microstructures with complex geometry at high resolution, produce fully enclosed internal cavities containing a solid or liquid, and use potentially any thermoplastic material without processing additives.

Mesoporous InN/In2O3 heterojunction with improved sensitivity and selectivity for room temperature NO2 gas sensing.

Establishing heterostructures is a good strategy to improve gas sensing performance, and has been studied extensively. In this work, mesoporous InN/In2O3 composite (InNOCs) heterostructures were prepared through a simple two-step strategy involving hydrothermal synthesis of In2O3 and subsequent nitriding into InN-composite In2O3 heterostructures. We found that the InN content has great influence on the resistance of InNOCs, and thus, the gas sensing performance. In particular, InNOC-36.9 (with InN content of 36.9% in the composites) shows an excellent sensing response towards different concentrations of NO2, as well as good stability after one week of exposure to 200 ppb NO2 at room temperature. The highest sensing response (ΔR/R0 ) is up to 1.8 for the low NO2 concentration of 5 ppb. Even more significantly, the theoretical limit of detection (LOD) of the InNOC-36.9 sensor is 31.7 ppt based on a signal-to-noise ratio of 3 (the measured LOD is 5 ppb), which is far below the US NAAQS  value (NO2: 53 ppb). In addition, a rational band structure model combined with a surface reaction model is proposed to explain the sensing mechanism.

Low Working-Temperature Acetone Vapor Sensor Based on Zinc Nitride and Oxide Hybrid Composites.

Transition-metal nitride and oxide composites are a significant class of emerging materials that have attracted great interest for their potential in combining the advantages of nitrides and oxides. Here, a novel class of gas sensing materials based on hybrid Zn3 N2 and ZnO composites is presented. The Zn3 N2 /ZnO (ZnNO) composites-based sensor exhibits selectivity and high sensitivity toward acetone vapor, and the sensitivity is dependent on the nitrogen content of the composites. The ZnNO-11.7 described herein possesses a low working temperature of 200 °C. The detection limit (0.07 ppm) is below the diabetes diagnosis threshold (1.8 ppm). In addition, the sensor shows high reproducibility and long-term stability.

Thermostabilization of inactivated polio vaccine in PLGA-based microspheres for pulsatile release.

Vaccines are a critical clinical tool in preventing illness and death due to infectious diseases and are regularly administered to children and adults across the globe. In order to obtain full protection from many vaccines, an individual needs to receive multiple doses over the course of months. However, vaccine administration in developing countries is limited by the difficulty in consistently delivering a second or third dose, and some vaccines, including the inactivated polio vaccine (IPV), must be injected more than once for efficacy. In addition, IPV does not remain stable over time at elevated temperatures, such as those it would encounter over time in the body if it were to be injected as a single-administration vaccine. In this manuscript, we describe microspheres composed of poly(lactic-co-glycolic acid) (PLGA) that can encapsulate IPV along with stabilizing excipients and release immunogenic IPV over the course of several weeks. Additionally, pH-sensitive, cationic dopants such as Eudragit E polymer caused clinically relevant amounts of stable IPV release upon degradation of the PLGA matrix. Specifically, IPV was released in two separate bursts, mimicking the delivery of two boluses approximately one month apart. In one of our top formulations, 1.4, 1.1, and 1.2 doses of the IPV serotype 1, 2, and 3, respectively, were released within the first few days from 50mg of particles. During the delayed, second burst, 0.5, 0.8, and 0.6 doses of each serotype, respectively, were released; thus, 50mg of these particles released approximately two clinical doses spaced a month apart. Immunization of rats with the leading microsphere formulation showed more robust and long-lasting humoral immune response compared to a single bolus injection and was statistically non-inferior from two bolus injections spaced 1 month apart. By minimizing the number of administrations of a vaccine, such as IPV, this technology can serve as a tool to aid in the eradication of polio and other infectious diseases for the improvement of global health.

Single-injection vaccines: Progress, challenges, and opportunities.

Currently, vaccination is the most efficient and cost-effective medical treatment for infectious diseases; however, each year 10 million infants remain underimmunized due to current vaccination schedules that require multiple doses to be administered across months or years. These dosing regimens are especially challenging in the developing world where limited healthcare access poses a major logistical barrier to immunization. Over the past four decades, researchers have attempted to overcome this issue by developing single-administration vaccines based on controlled-release antigen delivery systems. These systems can be administered once, but release antigen over an extended period of time to elicit both primary and secondary immune responses resulting in antigen-specific immunological memory. Unfortunately, unlike controlled release systems for drugs, single-administration vaccines have yet to be commercialized due to poor antigen stability and difficulty in obtaining unconventional release kinetics. This review discusses the current state of single-administration vaccination, challenges delaying the development of these vaccines, and potential strategies for overcoming these challenges.

Nano-structured ternary niobium titanium nitrides as durable non-carbon supports for oxygen reduction reaction.

A novel, highly stable, non-carbon support system for the oxygen reduction reaction (ORR) has been discovered in the form of nano-structured (NbxTi1-x)Ns (x = 0.25 and 0.5). Template-free, solid-solid separation synthetic approaches have been used for preparing these materials. The (NbxTi1-x)N materials have high specific surface area and better electronic conductivity than carbon-based supports.