The investigation of intracellular metabolism is the mainstay in the biotechnology and physiology settings. Intracellular metabolic rates are commonly evaluated using labeling pattern of the identified metabolites obtained from stable isotope labeling experiments. The labeling pattern or mass distribution vector describes the fractional abundances of all isotopologs with different masses as a result of isotopic labeling, which are typically resolved using mass spectrometry. Because naturally occurring isotopes and isotopic impurity also contribute to measured signals, the measured patterns must be corrected to obtain the labeling patterns. Since contaminant isotopologs with the same nominal mass can be resolved using modern mass spectrometers with high mass resolution, the correction process should be resolution dependent. Here we present a software tool, ElemCor, to perform correction of such data in a resolution-dependent manner. The tool is based on mass difference theory (MDT) and information from unlabeled samples (ULS) to account for resolution effects. MDT is a mathematical theory and only requires chemical formulae to perform correction. ULS is semi-empirical and requires additional measurement of isotopologs from unlabeled samples. We validate both methods and show their improvement in accuracy and comprehensiveness over existing methods using simulated data and experimental data from Saccharomyces cerevisiae. The tool is available at https://github.com/4dsoftware/elemcor . We present a software tool based on two methods, MDT and ULS, to correct LC-MS data from isotopic labeling experiments for natural abundance and isotopic impurity. We recommend MDT for low-mass compounds for cost efficiency in experiments, and ULS for high-mass compounds with relatively large spectral inaccuracy that can be tracked by unlabeled standards.

Although treatment with immune checkpoint inhibitors provides promising benefit for patients with cancer, optimal use is encumbered by high resistance rates and requires a thorough understanding of resistance mechanisms. We observed that tumors treated with PD-1/PD-L1 blocking antibodies develop resistance through the upregulation of CD38, which is induced by all-trans retinoic acid and IFNβ in the tumor microenvironment. In vitro and in vivo studies demonstrate that CD38 inhibits CD8+ T-cell function via adenosine receptor signaling and that CD38 or adenosine receptor blockade are effective strategies to overcome the resistance. Large data sets of human tumors reveal expression of CD38 in a subset of tumors with high levels of basal or treatment-induced T-cell infiltration, where immune checkpoint therapies are thought to be most effective. These findings provide a novel mechanism of acquired resistance to immune checkpoint therapy and an opportunity to expand their efficacy in cancer treatment. Significance: CD38 is a major mechanism of acquired resistance to PD-1/PD-L1 blockade, causing CD8+ T-cell suppression. Coinhibition of CD38 and PD-L1 improves antitumor immune response. Biomarker assessment in patient cohorts suggests that a combination strategy is applicable to a large percentage of patients in whom PD-1/PD-L1 blockade is currently indicated. Cancer Discov; 8(9); 1156–75. ©2018 AACR. See related commentary by Mittal et al., p. 1066. This article is highlighted in the In This Issue feature, p. 1047

Targeting oxidative phosphorylation overcomes ibrutinib resistance in mantle cell lymphoma.

The authors review the latest knowledge of amino acid metabolism in hematologic malignancies and the clinical relevance and potential of amino acid therapeutic targeting.

How cells coordinate cell cycling with cell survival and death remains incompletely understood. Here, we show that cell cycle arrest has a potent suppressive effect on ferroptosis, a form of regulated cell death induced by overwhelming lipid peroxidation at cellular membranes. Mechanistically, cell cycle arrest induces diacylglycerol acyltransferase (DGAT)–dependent lipid droplet formation to sequester excessive polyunsaturated fatty acids (PUFAs) that accumulate in arrested cells in triacylglycerols (TAGs), resulting in ferroptosis suppression. Consequently, DGAT inhibition orchestrates a reshuffling of PUFAs from TAGs to phospholipids and re-sensitizes arrested cells to ferroptosis. We show that some slow-cycling antimitotic drug–resistant cancer cells, such as 5-fluorouracil–resistant cells, have accumulation of lipid droplets and that combined treatment with ferroptosis inducers and DGAT inhibitors effectively suppresses the growth of 5-fluorouracil–resistant tumors by inducing ferroptosis. Together, these results reveal a role for cell cycle arrest in driving ferroptosis resistance and suggest a ferroptosis-inducing therapeutic strategy to target slow-cycling therapy-resistant cancers.

Key Points: We used molecular dynamics, saturation mutagenesis, and enzymologic screening to develop a glutaminase-free mutant (Q59L) l-ASP. We then used Q59L to show that glutaminase activity is not required for l-ASP activity against ASNS-negative cancer cells.