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Supplementary Figure 1. Synthesis and pharmacokinetic characterization of AG-221. Supplementary Figure 2. Biochemical attributes of AG-221. Supplementary Figure 3. Biochemistry of AG-221 with respect to substrate and cofactor. Supplementary Figure 4. TF-1 IDH2R140Q cells treated with AG-221. Supplementary Figure 5. AG-221 can reverse the block in EPO-induced differentiation caused by the expression of IDH2R140Q in the TF-1 erythroleukemia cell line. Supplementary Figure 6. Pharmacokinetics/pharmacodynamics of AG-221 in IDH2R140Qmutant U87MG xenograft tumor-bearing mice. Supplementary Figure 7. AG-221 does not affect intrinsic hematological parameters or body weight. Supplementary Figure 8. AG-221 strongly reduces the number of human IDH2R140Q blasts in the liver and spleen in AML-1, AML-2, and AML-3. Supplementary Figure 9. Flow cytometry analyses of bone marrow-derived hCD45+ cells in primary human AML xenograft models. Supplementary Figure 10. Affinity of the IDH2R140Q homodimer for NADPH. Supplementary Figure 11. Electron density map diagrams for bound ligands for IDH2R140Q co-complex structures determined by X-ray crystallography.

Supplementary Table 1. Drug metabolism and pharmacokinetic attributes of AG-221. Supplementary Table 2. Selectivity of AG-221 confirmed by testing against a panel of kinases. Supplementary Table 3. Clinical characteristics of patients with IDH2R140Q-mutated AML. Supplementary Table 4. Treated NSG mice (AML-1, AML-2, AML-3) engrafted with human IDH2R140Q mononuclear cells display stable levels of AG-221 in serum. Supplementary Table 5. AG-221 inhibits 2HG production in models AML-1, AML-2, and AML-3. Supplementary Table 6. Summary of pharmacokinetics/pharmacodynamics in primary human acute myeloid leukemia xenograft model (AML-4). Supplementary Table 7. Summary of data collection and refinement statistics. Supplementary Table 8. Percentage of human chimerism in peripheral blood in models AML-1 and AML-2.

Supplementary Methods

Purpose. Hepatocellular carcinoma (HCC), a malignant tumor that exists worldwide, has a high morbidity and mortality rate. Previous studies have reported that lncRNA NR2F1-AS1 plays a critical role in several cancers. Here, we aimed to investigate the biological function of NR2F1-AS1 and its molecular mechanism in the migration and invasion of HCC. Methods. Quantitative real-time PCR (qRT-PCR) was performed to analyze NR2F1-AS1 expression in HCC. The biological function was investigated by transwell invasion and migration assays. The protein level was identified by Western blot. In addition, the downstream targets of NR2F1-AS1 and miR-642a were confirmed by luciferase reporter assays. Results. NR2F1-AS1 was significantly upregulated in HCC and associated with the poor prognosis of HCC patients. Biological function experiments revealed that the silence of NR2F1-AS1 suppressed cell invasion and migration in HCC. More importantly, NR2F1-AS1 directly interacted with miR-642a and negatively regulated miR-642a. DEK was the target of miR-642a, and NR2F1-AS1 positively regulated DEK expression by suppressing miR-642a. Conclusion. Taken together, it is the first time we discovered the interaction of NR2F1-AS1 with miR-642a in modulating HCC cell invasion and migration.

Somatic gain-of-function mutations in isocitrate dehydrogenases (IDH) 1 and 2 are found in multiple hematologic and solid tumors, leading to accumulation of the oncometabolite (R)-2-hydroxyglutarate (2HG). 2HG competitively inhibits α-ketoglutarate–dependent dioxygenases, including histone demethylases and methylcytosine dioxygenases of the TET family, causing epigenetic dysregulation and a block in cellular differentiation. In vitro studies have provided proof of concept for mutant IDH inhibition as a therapeutic approach. We report the discovery and characterization of AG-221, an orally available, selective, potent inhibitor of the mutant IDH2 enzyme. AG-221 suppressed 2HG production and induced cellular differentiation in primary human IDH2 mutation–positive acute myeloid leukemia (AML) cells ex vivo and in xenograft mouse models. AG-221 also provided a statistically significant survival benefit in an aggressive IDH2R140Q-mutant AML xenograft mouse model. These findings supported initiation of the ongoing clinical trials of AG-221 in patients with IDH2 mutation–positive advanced hematologic malignancies. Significance: Mutations in IDH1/2 are identified in approximately 20% of patients with AML and contribute to leukemia via a block in hematopoietic cell differentiation. We have shown that the targeted inhibitor AG-221 suppresses the mutant IDH2 enzyme in multiple preclinical models and induces differentiation of malignant blasts, supporting its clinical development. Cancer Discov; 7(5); 478–93. ©2017 AACR. See related commentary by Thomas and Majeti, p. 459. See related article by Shih et al., p. 494. This article is highlighted in the In This Issue feature, p. 443

Pyruvate kinase (PK) deficiency is a rare genetic disease that causes chronic hemolytic anemia. There are currently no targeted therapies for PK deficiency. Here, we describe the identification and characterization of AG-348, an allosteric activator of PK that is currently in clinical trials for the treatment of PK deficiency. We demonstrate that AG-348 can increase the activity of wild-type and mutant PK enzymes in biochemical assays and in patient red blood cells treated ex vivo. These data illustrate the potential for AG-348 to restore the glycolytic pathway activity in patients with PK deficiency and ultimately lead to clinical benefit.

The North China Craton (NCC), one of the oldest continental blocks in Asia, has a complicated evolutionary history with the age of the old crust up to 3.8 Ga and records the important geological events of the Earth. The Sijiaying BIF, the largest banded iron formation located in eastern Hebei province, in the east part of NCC, records an important thermo-tectonic event; Previous studies focused on the geological description, but the timing and tectonic setting have been unclear so far. The Sijiaying BIF is hosted in Neoarchean metamorphic rocks, which includes biotite-leptynite, hornblende plagioclase-gneiss and biotite plagioclase-gneiss. Using major element contents and ratios of the host rocks, the protoliths of the hornblende plagioclase-gneiss and biotite plagioclase-gneiss are shown to be dacite, whereas those of the biotite-leptynites are sedimentary rocks. Based on geology and geochemistry of the host rocks, we infer that the Sijiaying BIF is an Algoma type deposit. SIMS zircon U–Pb dating shows: igneous zircons from the biotite plagioclase-gneiss and hornblende plagioclase-gneiss have U–Pb ages of 2535 ± 8 Ma and 2543 ± 14 Ma, respectively; zircons from the biotite-leptynite have an age of 2537 ± 13 Ma. We infer that the ages of 2543–2535 Ma represent the time of the Sijiaying BIF. PAAS-normalized REY profiles of the ore samples are characterized by LREE depletion and HREE enrichment, positive La and Eu anomalies and an average Y/Ho weight ratio of 32, indicating a mixture of submarine hydrothermal fluids and deep seawater. REE data was combined with the δ18O analyzed by previous researcher of the ore from individual magnetite bands to infer that the Sijiaying BIF precipitated from hydrothermal fluids discharging on the sea floor. The hornblende plagioclase-gneiss and biotite plagioclase-gneiss that are enriched in LILE and LREE and depleted in high field strength elements (HFSE) Nb, Ta and Ti, have geochemical signatures that are similar to those of island-arc volcanic rocks. The biotite-leptynite shows the characteristics of continental island-arc. Based on the geology and geochemistry of associated wall-rocks and ore samples, we suggest that the Sijiaying BIF formed in a back-arc basin in Neoarchean.

Scheelite (CaWO4), a hydrothermal mineral commonly displaying enrichment in Mo (up to 16%) and/or rare earth elements (REE), is the main economic mineral in the Gaojiabang porphyry-skarn type W-Mo deposit, East China. Based on microscopic observations and in-situ LA-ICP-MS and LA-MC-ICP-MS analysis, three groups of scheelite, each with different geochemical characteristics, can be recognized. This evidence provides a good basis for considering the behavior of some trace elements in scheelite and how they may constrain ore genesis in porphyry-skarn systems. P-group scheelite occurs inside the porphyry rocks in the form of vein-hosted or disseminated scheelite. These have the lowest (87Sr/86Sr)i values (0.7089–0.7108), lowest Mo concentration (mean 213 ppm; n = 51), LREE/HREE ratios of 16.33–84.91, and highest ΣREE concentration (191–405 ppm) with downwards-sloping REE fractionation trends. S-group scheelite occurs in skarns, coexists with skarn minerals (garnet, diopside, epidote, etc.) and has the highest (87Sr/86Sr)i values (0.7103–0.7113), highest Mo concentration (mean 1,323 ppm; n = 29), highest LREE/HREE ratio (62.6–164), and lowest ΣREE concentrations (68.5–112 ppm), also with strongly downwards-sloping REE fractionation trends. H-group scheelite occurs within veins hosted by hornfels and displays moderate (87Sr/86Sr)i (0.70104–0.70122), Mo (mean 806 ppm; n = 17), LREE/HREE ratio (16.32–42.55), and ΣREE concentration (98.3–167.6 ppm). During hydrothermal precipitation of scheelite, changing redox state plays a major role in controlling Mo behavior. Both the precipitation of early skarn minerals and changing redox states of ore-forming fluids likely result in an increase in Mo, and corresponding decrease of HREE, in scheelite (particularly in S-group scheelite). The Sr-isotope study indicates that crustal materials provided the main source for W-Mo-bearing ore-forming fluids. Furthermore, both fluid mixing and fluid-rock interaction played an important role in the evolution of ore-forming fluids: Formation water or groundwater was likely involved in ore formation. A geochemical model is described combining data for the different types of scheelite, and the behavior of Mo, REEs and Sr-isotopes, to constrain the evolution of ore-forming fluids and constrain ore genesis in the porphyry-skarn system at Gaojiabang. This study contributes to the fingerprinting of ore deposits using scheelite geochemistry.

AG-221 or enasidenib is a first-in-class selective inhibitor of mutated isocitrate dehydrogenase 2 (IDH2) with early demonstrated clinical efficacy in acute myeloid leukemia as a single agent, yet with persistence of mutant IDH2 clones. Two papers in this issue of Cancer Discovery provide further insight into the biological activity of AG-221 in promoting differentiation of IDH2 mutant cells and reversing aberrant DNA methylation over time, and demonstrating pre-clinical activity in combination with a targeted FLT3 kinase inhibitor to eliminate IDH2 mutant clones.

Enchondromas are benign cartilage tumors and precursors to malignant chondrosarcomas. Somatic mutations in the isocitrate dehydrogenase genes (IDH1 and IDH2) are present in the majority of these tumor types. How these mutations cause enchondromas is unclear. Here, we identified the spectrum of IDH mutations in human enchondromas and chondrosarcomas and studied their effects in mice. A broad range of mutations was identified, including the previously unreported IDH1-R132Q mutation. These mutations harbored enzymatic activity to catalyze α-ketoglutarate to d-2-hydroxyglutarate (d-2HG). Mice expressing Idh1-R132Q in one allele in cells expressing type 2 collagen showed a disordered growth plate, with persistence of type X-expressing chondrocytes. Chondrocyte cell cultures from these animals or controls showed that there was an increase in proliferation and expression of genes characteristic of hypertrophic chondrocytes with expression of Idh1-R132Q or 2HG treatment. Col2a1-Cre;Idh1-R132Q mutant knock-in mice (mutant allele expressed in chondrocytes) did not survive after the neonatal stage. Col2a1-Cre/ERT2;Idh1-R132 mutant conditional knock-in mice, in which Cre was induced by tamoxifen after weaning, developed multiple enchondroma-like lesions. Taken together, these data show that mutant IDH or d-2HG causes persistence of chondrocytes, giving rise to rests of growth-plate cells that persist in the bone as enchondromas.

Attenuation of millimeter waves induced by sand and dust storms along horizontal paths is analyzed and its expressions applied to earth-space paths is presented. Graphs of the specific attenuation and attenuation versus height and visibility are gotten for two kinds of particle size distribution, namely exponential, lognormal. Results obtained are briefly discussed.

Based on the measuring data of landforms, high-resolution seismic profiles, drilling cores, etc, a diapir body was found in the north of the modem Yellow River delta. The diapir body with a length of 5 km and a width of 1 km is distributed on the middle to low part of the slope of an abandoned delta lobe. Its formation is related to the deformity of the soft stratum which is deposited in the prodelta and covered by the stratum of the mouth bar sediments. Research results show that its formation is very different from the Mississippi River delta’s diapirs but related to the erosion of the seabed and occurs on the location with a large eroded quantity. The soft stratum and its diapir body can result in a great hazard to marine buildings.