The Transcription Factor Bhlhe40 Programs Mitochondrial Regulation of Resident CD8+ T Cell Fitness and Functionality.

Tissue-resident memory CD8+ T (Trm) cells share core residency gene programs with tumor-infiltrating lymphocytes (TILs). However, the transcriptional, metabolic, and epigenetic regulation of Trm cell and TIL development and function is largely undefined. Here, we found that the transcription factor Bhlhe40 was specifically required for Trm cell and TIL development and polyfunctionality. Local PD-1 signaling inhibited TIL Bhlhe40 expression, and Bhlhe40 was critical for TIL reinvigoration following anti-PD-L1 blockade. Mechanistically, Bhlhe40 sustained Trm cell and TIL mitochondrial fitness and a functional epigenetic state. Building on these findings, we identified an epigenetic and metabolic regimen that promoted Trm cell and TIL gene signatures associated with tissue residency and polyfunctionality. This regimen empowered the anti-tumor activity of CD8+ T cells and possessed therapeutic potential even at an advanced tumor stage in mouse models. Our results provide mechanistic insights into the local regulation of Trm cell and TIL function.

Overexpression of the energy metabolism transcriptome within clonal plasma cells is associated with the pathogenesis and outcomes of patients with multiple myeloma.

Altered energy metabolism and changes in glycolytic and oxidative phosphorylation pathways are hallmarks of all cancer cells. The expression of select genes associated with the production of various enzymes and proteins involved in glycolysis and oxidative phosphorylation were assessed in the clonal plasma cells derived from patients with newly diagnosed multiple myeloma (NDMM) enrolled in the Multiple Myeloma Research Foundation (MMRF) CoMMpass data set. A scoring system consisting of assigning a point for every gene where their fragments per kilobase of transcript per million (FPKM) was above the median yielded a minimum of 0 and a maximum of 12 for the set of genes in the glycolytic and oxidative phosphorylation pathways to create a total energy metabolism molecular signature (EMMS) score. This EMMS score was independently associated with worse progression free survival (PFS) and overall survival (OS) outcomes of patients with NDMM. A higher EMMS score was more likely to be present in clonal plasma cells derived from Multiple myeloma (MM) patients than those from patients with monoclonal gammopathy of undetermined significance (MGUS). This was functionally confirmed by the clonal plasma cells from MM patients having a higher rate of mitochondrial and glycolysis-derived ATP formation than clonal plasma cells from MGUS patients. Thus, this study provides evidence for the effect of energy metabolism within clonal plasma cells on pathogenesis and outcomes of patients with MM. Exploiting the energy-producing metabolic pathways within clonal plasma cells for diagnostic and therapeutic purposes in MM should be explored in the future.

Tyr phosphorylation of PDP1 toggles recruitment between ACAT1 and SIRT3 to regulate the pyruvate dehydrogenase complex.

Mitochondrial pyruvate dehydrogenase complex (PDC) is crucial for glucose homeostasis in mammalian cells. The current understanding of PDC regulation involves inhibitory serine phosphorylation of pyruvate dehydrogenase (PDH) by PDH kinase (PDK), whereas dephosphorylation of PDH by PDH phosphatase (PDP) activates PDC. Here, we report that lysine acetylation of PDHA1 and PDP1 is common in epidermal growth factor (EGF)-stimulated cells and diverse human cancer cells. K321 acetylation inhibits PDHA1 by recruiting PDK1, and K202 acetylation inhibits PDP1 by dissociating its substrate PDHA1, both of which are important in promoting glycolysis in cancer cells and consequent tumor growth. Moreover, we identified mitochondrial ACAT1 and SIRT3 as the upstream acetyltransferase and deacetylase, respectively, of PDHA1 and PDP1, while knockdown of ACAT1 attenuates tumor growth. Furthermore, Y381 phosphorylation of PDP1 dissociates SIRT3 and recruits ACAT1 to PDC. Together, hierarchical, distinct posttranslational modifications act in concert to control molecular composition of PDC and contribute to the Warburg effect.

Lysine acetylation activates 6-phosphogluconate dehydrogenase to promote tumor growth.

Although the oxidative pentose phosphate pathway is important for tumor growth, how 6-phosphogluconate dehydrogenase (6PGD) in this pathway is upregulated in human cancers is unknown. We found that 6PGD is commonly activated in EGF-stimulated cells and human cancer cells by lysine acetylation. Acetylation at K76 and K294 of 6PGD promotes NADP(+) binding to 6PGD and formation of active 6PGD dimers, respectively. Moreover, we identified DLAT and ACAT2 as upstream acetyltransferases of K76 and K294, respectively, and HDAC4 as the deacetylase of both sites. Expressing acetyl-deficient mutants of 6PGD in cancer cells significantly attenuated cell proliferation and tumor growth. This is due in part to reduced levels of 6PGD products ribulose-5-phosphate and NADPH, which led to reduced RNA and lipid biosynthesis as well as elevated ROS. Furthermore, 6PGD activity is upregulated with increased lysine acetylation in primary leukemia cells from human patients, providing mechanistic insights into 6PGD upregulation in cancer cells.

Relaxation of the Interface Resistance between Solid Electrolyte and 5 V-Class Positive Electrode.

Solid-state Li batteries using 5 V-class positive electrode materials display a higher energy density. However, the high resistance at the interface of the electrolyte and positive electrode (interface resistance, Ri) hinders their practical applications. Here, we report the relaxation of Ri between a solid electrolyte (Li3PO4) and a 5 V-class electrode (LiCo0.5Mn1.5O4). Although Ri is small at the Mn3+/4+ redox voltage of 4.0 V vs Li/Li+ (11 Ω cm2), it rapidly increases by more than 2 orders of magnitude as the voltage increases above the Co3+/4+ redox voltage of 5.2 V vs Li/Li+. After the applied voltage is reduced to 4.0 V vs Li/Li+, Ri decays to the original value after 3 h. The relaxation of Ri after exposure to high voltages suggests that the increase in Ri above 5 V vs Li/Li+ is attributable to the formation of an interfacial layer at the LPO/LCMO interface.

Ionic Rectification across Ionic and Mixed Conductor Interfaces.

In electrochemical devices, it is important to control the ionic transport between the electrodes and solid electrolytes. However, it is difficult to tune the transport without applying an electric field. This paper presents a method to modulate the transport via tuning of the electrochemical potential difference by controlling the electronic states at the interfaces. We fabricated thin-film solid-state Li batteries using LiTi2O4 thin films as positive electrodes. The spontaneous Li-ion transport between the solid electrolyte and LiTi2O4 is controlled by tuning the electrochemical potential difference via use of an electrically conducting Nb-doped SrTiO3 substrate. This study establishes the foundation for rectifying the ionic transport via electronic energy band alignment.

High Li-Ion Conductivity in Li{N(SO2F)2}(NCCH2CH2CN)2 Molecular Crystal.

There is an urgent need to develop solid electrolytes based on organic molecular crystals for application in energy devices. However, the quest for molecular crystals with high Li-ion conductivity is still in its infancy. In this study, the high Li-ion conductivity of a Li{N(SO2F)2}(NCCH2CH2CN)2 molecular crystal is reported. The crystal shows a Li-ion conductivity of 1 × 10-4 S cm-1 at 30 °C and 1 × 10-5 S cm-1 at -20 °C, with a low activation energy of 28 kJ mol-1. The conductivity at 30 °C is one of the highest values attainable by molecular crystals, whereas that at -20 °C is approximately 2 orders of magnitude higher than previously reported values. Furthermore, the all-solid-state Li-battery fabricated using this solid electrolyte demonstrates stable cycling, thereby maintaining 90% of the initial capacity after 100 charge-discharge cycles. The finding of high Li-ion conductivity in molecular crystals paves the way for their application in all-solid-state Li-batteries.

Interfacial Superconductivity in FeSe Ultrathin Films on SrTiO_{3} Probed by In Situ Independently Driven Four-Point-Probe Measurements.

We investigated the superconducting transport properties of the one-unit-cell FeSe ultrathin films epitaxially grown on undoped SrTiO_{3}(001) (STO) with a well-defined surface structure by in situ independently-driven four-point-probe measurements. Our results unambiguously revealed the detection of the two-dimensional electrical conduction of the films without parallel conduction through the underlying substrate, both in the normal and superconducting states. The monolayer film exhibited a superconducting transition at an onset temperature of 40 K. Surprisingly, the onset of superconductivity was constantly observed at 40 K even for three- and five-unit-cell-thick FeSe films, even though the normal resistivity decreased with increasing thickness. These results agree with the picture of the interfacial superconductivity, where only the FeSe/STO interface and/or the adjacent first layer of FeSe becomes superconducting while the upper layers stay in the normal metallic state. The observed T_{c} is much lower than that reported by a previous in situ transport measurement for FeSe/Nb:STO but consistent with the results obtained by ex situ measurements for FeSe-undoped STO with a capping layer. This suggests that the capping layer is not an essential factor to limit T_{c}. We rather propose that the charge transfer from the doped substrate has a key role to achieve the higher temperature superconductivity in the one-unit-cell FeSe.

Epitaxial Growth of Single-Phase Magnesium Dihydride Thin Films.

We describe the epitaxial growth process of single-phase magnesium dihydride (MgH2) thin films on MgO(100) substrates, achieved by reactive magnetron sputtering. We find that direct growth at substrate temperatures higher than 100 °C leads to partial MgH2 decomposition to Mg, hindering single-phase epitaxy of MgH2. To improve the crystallinity and suppress the decomposition of Mg, we optimize MgH2 growth using a two-step process, consisting of (1) precursor growth at room temperature and (2) postdeposition annealing at 380 °C, under a pressure of 1.0 × 105 Pa with H2 (4%)/Ar (96%) premixed gases. Using this two-step process, we obtain single-phase MgH2 epitaxial films with high crystallinity, transparency, and resistivity. Further, the application of this method to grow MgH2 thin films on different MgF2 and Al2O3 substrates enables us to use the epitaxial effects to control the growth orientation of MgH2 thin films; we show that MgH2(100) and MgH2(001) epitaxial thin films can be grown on Al2O3(001) and MgF2(001) substrates, respectively.

Tyrosine phosphorylation of mitochondrial pyruvate dehydrogenase kinase 1 is important for cancer metabolism.

Many tumor cells rely on aerobic glycolysis instead of oxidative phosphorylation for their continued proliferation and survival. Myc and HIF-1 are believed to promote such a metabolic switch by, in part, upregulating gene expression of pyruvate dehydrogenase (PDH) kinase 1 (PDHK1), which phosphorylates and inactivates mitochondrial PDH and consequently pyruvate dehydrogenase complex (PDC). Here we report that tyrosine phosphorylation enhances PDHK1 kinase activity by promoting ATP and PDC binding. Functional PDC can form in mitochondria outside of the matrix in some cancer cells and PDHK1 is commonly tyrosine phosphorylated in human cancers by diverse oncogenic tyrosine kinases localized to different mitochondrial compartments. Expression of phosphorylation-deficient, catalytic hypomorph PDHK1 mutants in cancer cells leads to decreased cell proliferation under hypoxia and increased oxidative phosphorylation with enhanced mitochondrial utilization of pyruvate and reduced tumor growth in xenograft nude mice. Together, tyrosine phosphorylation activates PDHK1 to promote the Warburg effect and tumor growth.

Atomic-scale visualization of oxide thin-film surfaces.

The interfaces of complex oxide heterostructures exhibit intriguing phenomena not observed in their constituent materials. The oxide thin-film growth of such heterostructures has been successfully controlled with unit-cell precision; however, atomic-scale understandings of oxide thin-film surfaces and interfaces have remained insufficient. We examined, with atomic precision, the surface and electronic structures of oxide thin films and their growth processes using low-temperature scanning tunneling microscopy. Our results reveal that oxide thin-film surface structures are complicated in contrast to the general perception and that atomically ordered surfaces can be achieved with careful attention to the surface preparation. Such atomically ordered oxide thin-film surfaces offer great opportunities not only for investigating the microscopic origins of interfacial phenomena but also for exploring new surface phenomena and for studying the electronic states of complex oxides that are inaccessible using bulk samples.

Quasiparticle Interference on Cubic Perovskite Oxide Surfaces.

We report the observation of coherent surface states on cubic perovskite oxide SrVO_{3}(001) thin films through spectroscopic-imaging scanning tunneling microscopy. A direct link between the observed quasiparticle interference patterns and the formation of a d_{xy}-derived surface state is supported by first-principles calculations. We show that the apical oxygens on the topmost VO_{2} plane play a critical role in controlling the coherent surface state via modulating orbital state.

Negligible "negative space-charge layer effects" at oxide-electrolyte/electrode interfaces of thin-film batteries.

In this paper, we report the surprisingly low electrolyte/electrode interface resistance of 8.6 Ω cm(2) observed in thin-film batteries. This value is an order of magnitude smaller than that presented in previous reports on all-solid-state lithium batteries. The value is also smaller than that found in a liquid electrolyte-based batteries. The low interface resistance indicates that the negative space-charge layer effects at the Li3PO(4-x)N(x)/LiCoO2 interface are negligible and demonstrates that it is possible to fabricate all-solid state batteries with faster charging/discharging properties.

Tyr-94 phosphorylation inhibits pyruvate dehydrogenase phosphatase 1 and promotes tumor growth.

Many cancer cells rely more on aerobic glycolysis (the Warburg effect) than mitochondrial oxidative phosphorylation and catabolize glucose at a high rate. Such a metabolic switch is suggested to be due in part to functional attenuation of mitochondria in cancer cells. However, how oncogenic signals attenuate mitochondrial function and promote the switch to glycolysis remains unclear. We previously reported that tyrosine phosphorylation activates and inhibits mitochondrial pyruvate dehydrogenase kinase (PDK) and phosphatase (PDP), respectively, leading to enhanced inhibitory serine phosphorylation of pyruvate dehydrogenase (PDH) and consequently inhibition of pyruvate dehydrogenase complex (PDC) in cancer cells. In particular, Tyr-381 phosphorylation of PDP1 dissociates deacetylase SIRT3 and recruits acetyltransferase ACAT1 to PDC, resulting in increased inhibitory lysine acetylation of PDHA1 and PDP1. Here we report that phosphorylation at another tyrosine residue, Tyr-94, inhibits PDP1 by reducing the binding ability of PDP1 to lipoic acid, which is covalently attached to the L2 domain of dihydrolipoyl acetyltransferase (E2) to recruit PDP1 to PDC. We found that multiple oncogenic tyrosine kinases directly phosphorylated PDP1 at Tyr-94, and Tyr-94 phosphorylation of PDP1 was common in diverse human cancer cells and primary leukemia cells from patients. Moreover, expression of a phosphorylation-deficient PDP1 Y94F mutant in cancer cells resulted in increased oxidative phosphorylation, decreased cell proliferation under hypoxia, and reduced tumor growth in mice. Together, our findings suggest that phosphorylation at different tyrosine residues inhibits PDP1 through independent mechanisms, which act in concert to regulate PDC activity and promote the Warburg effect.

Charge-density wave in Ca-intercalated bilayer graphene induced by commensurate lattice matching.

We report the emergence of a charge-density wave (CDW) in Ca-intercalated bilayer graphene (C_{6}CaC_{6}), the thinnest limit of superconducting C_{6}Ca, observed by low-temperature, high-magnetic-field scanning tunneling microscopy or spectroscopy, and angle-resolved photoemission spectroscopy. While the possible superconductivity was not observed in epitaxially grown C_{6}CaC_{6} on a SiC substrate, a CDW order different from that observed on the surface of bulk C_{6}Ca was observed. It is inferred that the CDW state is induced by the potential modulation due to the commensurate lattice matching between the C_{6}CaC_{6} film and the SiC substrate.

Ca intercalated bilayer graphene as a thinnest limit of superconducting C6Ca.

Success in isolating a 2D graphene sheet from bulky graphite has triggered intensive studies of its physical properties as well as its application in devices. Graphite intercalation compounds (GICs) have provided a platform of exotic quantum phenomena such as superconductivity, but it is unclear whether such intercalation is feasible in the thinnest 2D limit (i.e., bilayer graphene). Here we report a unique experimental realization of 2D GIC, by fabricating calcium-intercalated bilayer graphene C(6)CaC(6) on silicon carbide. We have investigated the structure and electronic states by scanning tunneling microscopy and angle-resolved photoemission spectroscopy. We observed a free-electron-like interlayer band at the Brillouin-zone center, which is thought to be responsible for the superconductivity in 3D GICs, in addition to a large π* Fermi surface at the zone boundary. The present success in fabricating Ca-intercalated bilayer graphene would open a promising route to search for other 2D superconductors as well as to explore its application in devices.

Imaging the evolution of d states at a strontium titanate surface.

Oxide electronics is a promising alternative to the conventional silicon-based semiconductor technology, owing to the rich functionalities of oxide thin films and heterostructures. In contrast to the silicon surface, however, the electronic structure of the SrTiO3 surface, the most important substrate for oxide thin films growth, is not yet completely understood. Here we report on the electronic states of a reconstructed (001) surface of SrTiO3 determined in real space, with scanning tunneling microscopy/spectroscopy and density functional theory calculations. We found a remarkable energy dependence of the spectroscopic image: Theoretical analysis reveals that symmetry breaking at the surface lifts the degeneracy in the t2g state (dxy, dyz, and dzx) of Ti 3d orbitals, whose anisotropic spatial distribution leads to a sharp transition in the spectroscopic image as a function of energy. The knowledge obtained here could be used to gain further insights into emergent phenomena at the surfaces and interfaces with SrTiO3.

Scanning tunneling microscopy/spectroscopy on perovskite oxide thin films deposited in situ.

Complex oxide surfaces and interfaces, consisting of two or more cations and oxygen anions, have attracted a great deal of attention because their properties are crucial factors in the performance of catalysts, fuel cells, and Li-ion batteries. However, atomic-scale investigations of these oxide surfaces have been hindered because of the difficulties in surface preparation. Here, we demonstrate atomic-scale surface studies of complex perovskite oxides and the initial growth processes in oxide epitaxial films deposited on (✓13 × âœ“13)-R33.7° reconstructed SrTiO3 (001) substrates using a scanning tunneling microscope integrated with a pulsed laser deposition system. The atomically ordered, reconstructed SrTiO3 (001) surface is stable under the typical conditions necessary for the growth of oxide thin films, and hence is considered suitable for the study of the initial growth processes in oxide films. The atomic-scale microscopic/spectroscopic characterizations performed here shed light on the microscopic origin of electronic properties observed in complex oxides and their heterostructures.

Structural Basis for Substrate Binding, Catalysis and Inhibition of Breast Cancer Target Mitochondrial Creatine Kinase by Covalent Inhibitor via Cryo-EM.

Mitochondrial creatine kinases are key players in maintaining energy homeostasis in cells by working in conjunction with cytosolic creatine kinases for energy transport from mitochondria to cytoplasm. High levels of MtCK observed in Her2+ breast cancer and inhibition of breast cancer cell growth by substrate analog, cyclocreatine, indicate dependence of cancer cells on the 'energy shuttle' for cell growth and survival. Hence, understanding the key mechanistic features of creatine kinases and their inhibition plays an important role in the development of cancer therapeutics. Herein, we present the mutational and structural investigation on understudied ubiquitous mitochondrial creatine kinase (uMtCK). Our cryo-EM structures and biochemical data on uMtCK showed closure of the loop comprising residue His61 is specific to and relies on creatine binding and the reaction mechanism of phosphoryl transfer depends on electrostatics in the active site. In addition, the previously identified covalent inhibitor CKi showed inhibition in breast cancer BT474 cells, however our biochemical and structural data indicated that CKi is not a potent inhibitor for breast cancer due to strong dependency on the covalent link formation and inability to induce conformational changes upon binding.