Search

RNA-seq and ATAC-seq of TRAF3-normal and -deficient B16F10 cells treated with vehicle control or IFNγ

Immune checkpoint blockade (ICB) therapy revolutionized cancer treatment, but many patients with impaired MHC-I expression remain refractory. Here, we combined FACS-based genome-wide CRISPR screens with a data-mining approach to identify drugs that can upregulate MHC-I without inducing PD-L1. CRISPR screening identified TRAF3, a suppressor of the NFκB pathway, as a negative regulator of MHC-I but not PD-L1. The Traf3-knockout gene expression signature is associated with better survival in ICB-naïve patients with cancer and better ICB response. We then screened for drugs with similar transcriptional effects as this signature and identified Second Mitochondria-derived Activator of Caspase (SMAC) mimetics. We experimentally validated that the SMAC mimetic birinapant upregulates MHC-I, sensitizes cancer cells to T cell–dependent killing, and adds to ICB efficacy. Our findings provide preclinical rationale for treating tumors expressing low MHC-I expression with SMAC mimetics to enhance sensitivity to immunotherapy. The approach used in this study can be generalized to identify other drugs that enhance immunotherapy efficacy.Significance:MHC-I loss or downregulation in cancer cells is a major mechanism of resistance to T cell–based immunotherapies. Our study reveals that birinapant may be used for patients with low baseline MHC-I to enhance ICB response. This represents promising immunotherapy opportunities given the biosafety profile of birinapant from multiple clinical trials.This article is highlighted in the In This Issue feature, p. 1307

Supplementary Figures S1-S8

The spin-dependent polaron dynamics in organic ferromagnets under driven electric fields are investigated by using the extended Su-Schrieffer-Heeger (SSH) model coupled with a nonadiabatic dynamics method. It is found that the spin-down polaron with the same spin orientation as the radicals drifts faster than the spin-up one under the same driven electric field. In an applicable range of driven electric fields, the velocity of the spin-down polaron is about 3.4 times that of the spin-up one. The dynamical property of the polaron with each spin (up or down) is asymmetric upon the reversal of the driven electric fields. The diverse dynamical properties of polarons with specific spins can be attributed to the spin nondegenerate polaron energy levels, the dipole moment generated by the asymmetrical polaron charge distributions and the strong electron-lattice coupling in organic ferromagnets. Our findings are expected to be useful for improving organic ferromagnet based spintronic devices.

PD-1 expression marks activated T cells susceptible to PD-1–mediated inhibition but not whether a PD-1–mediated signal is being delivered. Molecular predictors of response to PD-1 immune checkpoint blockade (ICB) are needed. We describe a monoclonal antibody (mAb) that detects PD-1 signaling through the detection of phosphorylation of the immunotyrosine switch motif (ITSM) in the intracellular tail of mouse and human PD-1 (phospho–PD-1). We showed PD-1+ tumor-infiltrating lymphocytes (TILs) in MC38 murine tumors had high phosphorylated PD-1, particularly in PD-1+TIM-3+ TILs. Upon PD-1 blockade, PD-1 phosphorylation was decreased in CD8+ TILs. Phospho–PD-1 increased in T cells from healthy human donors after PD-1 engagement and decreased in patients with Hodgkin lymphoma following ICB. These data demonstrate that phosphorylation of the ITSM motif of PD-1 marks dysfunctional T cells that may be rescued with PD-1 blockade. Detection of phospho–PD-1 in TILs is a potential biomarker for PD-1 immunotherapy responses.

Immune checkpoint blockade (ICB) therapy revolutionized cancer treatment, but many patients with impaired MHC-I expression remain refractory. Here, we combined FACS-based genome-wide CRISPR screens with a data-mining approach to identify drugs that can upregulate MHC-I without inducing PD-L1. CRISPR screening identified TRAF3, a suppressor of the NFκB pathway, as a negative regulator of MHC-I but not PD-L1. The Traf3-knockout gene expression signature is associated with better survival in ICB-naïve patients with cancer and better ICB response. We then screened for drugs with similar transcriptional effects as this signature and identified Second Mitochondria-derived Activator of Caspase (SMAC) mimetics. We experimentally validated that the SMAC mimetic birinapant upregulates MHC-I, sensitizes cancer cells to T cell–dependent killing, and adds to ICB efficacy. Our findings provide preclinical rationale for treating tumors expressing low MHC-I expression with SMAC mimetics to enhance sensitivity to immunotherapy. The approach used in this study can be generalized to identify other drugs that enhance immunotherapy efficacy. Significance: MHC-I loss or downregulation in cancer cells is a major mechanism of resistance to T cell–based immunotherapies. Our study reveals that birinapant may be used for patients with low baseline MHC-I to enhance ICB response. This represents promising immunotherapy opportunities given the biosafety profile of birinapant from multiple clinical trials. This article is highlighted in the In This Issue feature, p. 1307

Immunotherapies that target programmed cell death protein 1 (PD-1) and its ligand PD-L1 as well as cytotoxic T-lymphocyte-associated protein 4 (CTLA4) have shown impressive clinical outcomes for multiple tumours. However, only a subset of patients achieves durable responses, suggesting that the mechanisms of the immune checkpoint pathways are not completely understood. Here, we report that PD-L1 translocates from the plasma membrane into the nucleus through interactions with components of the endocytosis and nucleocytoplasmic transport pathways, regulated by p300-mediated acetylation and HDAC2-dependent deacetylation of PD-L1. Moreover, PD-L1 deficiency leads to compromised expression of multiple immune-response-related genes. Genetically or pharmacologically modulating PD-L1 acetylation blocks its nuclear translocation, reprograms the expression of immune-response-related genes and, as a consequence, enhances the anti-tumour response to PD-1 blockade. Thus, our results reveal an acetylation-dependent regulation of PD-L1 nuclear localization that governs immune-response gene expression, and thereby advocate targeting PD-L1 translocation to enhance the efficacy of PD-1/PD-L1 blockade.

Adult T-cell leukemia/lymphoma (ATLL) in Japan presents at a median age of 70 years and only 5% of patients are

BackgroundCancer immunotherapy, especially immune checkpoint blockade (ICB) therapy, is leading to a paradigm shift in cancer treatment, as a small percentage of cancer patients have obtained durable remission following ICB treatment. Successful ICB responses rely on cancer cells presenting antigens to the cell surface via the major histocompatibility complex (MHC), which activates antigen-specific T-lymphocytes to kill cancer cells. Type-I MHC (MHC-I) is wildly expressed in all cell types and mediates the interaction with cytotoxic CD8 T cells. However, over 65% of cancer patients are estimated to show defects in MHC-I-mediated antigen presentation, including downregulation of its expression that can lead to primary or acquired resistance to ICB therapy, and therapeutic strategies to effectively restore or boost MHC-I are limited.MethodsHere, we employed a CRISPR screening approach with dual-marker FACS sorting to identify factors that decouple the regulation of MHC-I and PD-L1. The experimentally validated target was used to generate a KO differential expression signature. Using this signature, we analyzed transcriptome data from drug perturbation studies to identify drugs that regulate MHC-I but not PD-L1. Finally, we validated the effect of the identified drug to enhance ICB response in a T-cell-dependent manner in vivo.ResultsCRISPR screens identified TRAF3, a suppressor of the NF-κB pathway, as a negative regulator of MHC-I but not PD-L1. The Traf3-knockout (Traf3-KO) gene expression signature is associated with better survival in ICB-naive cancer patients and better ICB response. We then screened for drugs with similar transcriptional effects as this signature and identified SMAC mimetics. We experimentally validated that the SMAC mimetic birinapant upregulates MHC-I, sensitizes cancer cells to T-cell-dependent killing, and adds to ICB efficacy. However, in cancer cells with high NF-κB activity, the effect of birinapant on MHC-I is weak, indicating context-dependent MHC-I regulation.ConclusionsIn summary, Traf3 deletion specifically upregulates MHC-I without inducing PD-L1 in response to various cytokines and sensitizes cancer cells to T-cell-driven cytotoxicity. The SMAC mimetic birinapant phenocopies Traf3-knockout and sensitizes MHC-I-low melanoma to ICB therapy. Further studies are needed to elucidate the context-dependencies of MHC-I regulation. Our findings provide preclinical rationale for treating some tumors expressing low MHC-I with SMAC mimetics to enhance sensitivity to immunotherapy. The approach used in this study can be generalized to identify other drugs that enhance immunotherapy efficacy.AcknowledgementsThis study was supported by grants from the NIH (R01CA234018 to XSL, R01AI137337 to BEG, P50CA101942-12 and P50CA206963 to GJF), Breast Cancer Research Foundation (BCRF-19-100 to XSL), Burroughs Wellcome Career Award in Medical Sciences (to BEG), and Sara Elizabeth O'Brien Trust Fellowship (to SG).We thank Drs. Kai Wucherpfennig and Deng Pan for their insightful suggestions on this study.Ethics ApprovalAll mice were housed in standard cage in Dana-Farber Cancer Institute Animal Resources Facility (ARF). All animal procedures were carried out under the ARF Institutional Animal Care and Use Committee (IACUC) protocol and were in accordance with the IACUC standards for the welfare of animals.

lsa(E) is a pleuromutilin, lincosamide, and streptogramin A (PLSA phenotype) resistance gene that was first described in S. aureus and was thought to have been transferred from Enterococcus sp. This study aimed to elucidate the prevalence of the lsa(E) gene among E. faecium isolates at a tertiary teaching hospital and to evaluate the transferability of the lsa(E) gene from E. faecium to S. aureus in vitro. A total of 96 E. faecium strains isolated from one hospital in Beijing in 2013 were analysed for quinupristin-dalfopristin (QDA) resistance genes, and multilocus sequence typing (MLST) was performed. The transferability of QDA resistance between ten E. faecium strains and four S. aureus strains was determined by filter mating. Genome sequencing of the transconjugant was performed. A total of 46 E. faecium isolates (46/96, 47.92%) tested positive for lsa(E), while two isolates (2/96, 2.08%) tested positive for lsa(A). Thirty-six lsa(E)-positive strains (36/46, 78.3%) belonged to ST78. Among 40 mating tests, lsa(E) was successfully transferred through one conjugation at a frequency of 1.125 × 10–7 transconjugants per donor. The QDA resistance of the transconjugant N7435-R3645 was expressed at a higher level (MIC = 16 mg/L) than that of the parent S. aureus strain (MIC = 0.38 mg/L). Next-generation sequencing (NGS) analysis of the transconjugant N7435-R3645 showed that the complete sequence of the lsa(E)-carrying plasmid pN7435-R3645 had a size of 92,396 bp and a G + C content of 33% (accession no. MT022086). The genetic map of pN7435-R3645 had high nucleotide similarity and shared the main open reading frame (ORF) features with two plasmids: E. faecium pMG1 (AB206333.1) and E. faecium LS170308 (CP025078.1). The rep gene of pN7435-R3645 showed 100% identity with that of pMG1, although it did not belong to the rep1-19 family but instead a unique rep family. Multiple antibiotic resistance genes, including lsa(E), aadE and lnu(B), erm(B), ant6-Ia, and lnu(B), were present on the plasmid. In conclusion, an lsa(E)-carrying plasmid that can be transferred by conjugation from E. faecium to S. aureus in vitro was identified. This multidrug resistance (MDR) pMG1-like plasmid may act as a vector in the dissemination of antimicrobial resistance among species.

In this work, the density functional theory (DFT) calculations are employed to investigate the effect of in-plane strain on the electronic and magnetic anisotropy of SrTiO-terminated SrTiO3 (110) surface. The two-dimensional electron gas (2DEG) at the surface comes from the surface Ti 3d orbitals and exhibits anisotropic transport properties. The ordering of the lowest conduction band states can be tuned by in-plane strain. In addition, the in-plane strain can effectively modify and reduce the electron effective mass of the SrTiO3 (110) surface, and the effect of the in-plane strain on the two directions of [001] and [ 1 1 ¯ 0 ] is different. The magnetic moment of the unstrained surface Ti atom is 0.342 μB. The magnetic moment increases monotonically with the increasing of in-plane tensile strain and decreases quickly with increasing of the in-plane compressive strain. Importantly, the in-plane strain can change the magnetic moment of the system as well as its magnetic anisotropy energy and direction of easy magnetization axis. In-plane tensile strain causes a switch of the magnetic easy axis between parallel and perpendicular to the STO (110) surface.

Providing a reliable numerical tool for the prediction of parametric roll and pure loss of stability is a difficult task. It is acknowledged that the occurrence of these two failure modes is mainly attributed to the variation of roll restoring arm, which should be calculated accurately. This work studies exclusively the roll restoring arm variation numerically and experimentally. First, partially restrained experimental tests at different constant heeling angles (up to 30°) are conducted to measure the variation of roll restoring arm in regular waves. Second, a numerical method for the calculation of roll restoring arm in waves taking radiation and diffraction moments into account is adopted. The calculation results are benchmarked against experimental model test data. Third, the characteristics of roll restoring arm variation and the effects of radiation and diffraction moments on roll restoring arm variation (GZRD) in head and following seas are investigated. Finally, the feasibility of GZ calculated by the integral of instantaneously average wetted surface is studied by comparing the results with that calculated by the integral of the fully instantaneously wetted surface. The research show that the GZRD can be ignored in following seas. The GZ in head seas consists of two important harmonic components, and GZRD is the main contribution of the second harmonic component. Therefore, GZRD should be calculated with consideration of the instantaneous heeling angles and Froude number in head seas, especially for the prediction of large amplitude roll motion at high speed. 1. Introduction Currently, the second-generation intact stability criteria are under development by the International Maritime Organization (IMO) (IMO SLF 53/WP.1-Add.1 2011; IMO SDC 3/WP.5 2016). Five different kinds of potentially dangerous dynamic stability failure modes in waves were proposed. Parametric roll and pure loss of stability are two of the stability failure modes that may lead to capsizing of a ship. According to the proposed drafts, a multilevel structure was adopted to make sure that numerical assessment procedures are only applied when the vulnerability to dynamical stability failures is beyond reasonable doubt. Assessment of the vulnerability criteria is normally based on simplified geometry characteristics or empirical formulas. However, direct stability assessment requires highly accurate prediction (Kobylinski 2012).

A reliable numerical tool for the prediction of the parametric roll is important in the development of the second generation intact stability criteria. The main difficulty of the prediction of the parametric roll comes from the significant coupling of the roll with other motions, especially the heave and pitch motions. In this paper, the numerical method for determining the parametric roll is studied and the results are benchmarked against the model test data. Firstly, a 3 degrees-of-freedom (DOF) time domain hybrid panel method is validated by experimental data in regular oblique waves and irregular head waves. Secondly, the verified 3DOF method is extended to a fully coupled 6DOF method with the course control. Finally, taking the C11 containership as the test case, the parametric roll in oblique waves is computed, and the effects of the surge, sway and yaw motions on the parametric roll are evaluated with the developed mathematical models. The results show that the effects of the surge and sway motions on the parametric roll is insignificant, while the effects of the yaw motion are more notable. The results of the roll amplitudes and the standard deviation of the roll angles show that the model based on 3DOF gives relatively conservative results, and can be directly used for the stability assessment of the parametric roll.

Parametric roll is an important stability failure mode in the second generation intact stability criteria, which are currently under development by International Maritime Organization (IMO). The roll restoring arm, which is one of the dominating factors for the prediction of parametric roll, should be calculated preciously. In this paper, firstly, the effects of the radiation and diffraction forces on the roll restoring arm (GZR&D) under different heeling angles and wave amplitudes are studied based on the body exact method, in which all the hydrodynamic forces are calculated by integrating wave pressure up to the wave surface. Secondly, the effects of radiation and diffraction forces on parametric roll are studied by the direct simulation using a coupled heave-roll-pitch model based on the three dimensional mixed source method. Finally, by taking one passenger ship as an example, the calculation accuracy of the body exact method is compared with the commonly used body linear method. The results show that the effect of radiation and diffraction forces on righting arm variation has nonlinear relationships with heeling angles and wave amplitudes, and the body exact method can provide more accurate prediction results for parametric roll in head waves than the body linear method. Therefore, the nonlinear effects of instantaneous roll angles and wave amplitudes on GZR&D variation in waves should be considered for the prediction of parametric roll in head waves.

Half-metallic materials have gained a lot of attention because of their unique properties and applications in spintronic devices. Despite the fact that these materials have been studied by several research groups there are very limited studies on their heterostructure (HS) systems. In the current study we have investigated the electronic and magnetic properties of (LaAlO3)6.5/(SrTiO3)2.5(111) HS using density functional theory (DFT) calculations. We demonstrate that the system exhibits a 100% spin-polarized two-dimensional electron gas (2DEG) which is extremely confined to the Ti 3d orbitals of the SrTiO3 layers. In particular, this system can keep its half-metallic properties under different in-plane strains from −3 to 2%. This property proves that this material has relatively stable half-metallic properties. In addition, the conducting and magnetic ground states of the system can also be tailored by changing in-plane strain and interfacial cation intermixing of La and Sr (Sr ⇔ La intermixing). By increasing the in-plane lattice parameters, this system has the ability to evolve from a nonmagnetic to a ferromagnetic metal and then to a half-metal and by further increasing the in-plane lattice parameter it becomes a ferromagnetic insulator. Sr ⇔ La intermixing can destroy the original half-metallic properties and the system exhibits an AFM Mott-type insulator phase. Our results demonstrate that the system has high potential for application in the field of spintronics, and opens the prospect of using LaAlO3/SrTiO3(111) HSs to explore quantum phase transitions.

Targeting immune checkpoint pathways, such as programmed death ligand-1 (PD-L1, also known as CD274 or B7-H1) or its receptor programmed cell death-1 (PD-1) has shown improved survival for patients with numerous types of cancers, not limited to lung cancer, melanoma, renal cell carcinoma, and Hodgkin lymphoma. PD-L1 is a co-inhibitory molecule whose expression on the surface of tumor cells is associated with worse prognosis in many tumors. Here we describe a splice variant (secPD-L1) that does not splice into the transmembrane domain, but instead produces a secreted form of PD-L1 that has a unique 18 amino acid tail containing a cysteine that allows it to homodimerize and more effectively inhibit lymphocyte function than monomeric soluble PD-L1. We show that recombinant secPD-L1 can dimerize and inhibit T-cell proliferation and IFN-gamma production in vitro. The secPD-L1 variant is expressed by malignant cells in vitro that also express high levels of full-length PD-L1. Transcriptomic analysis of gene expression across The Cancer Genome Atlas found the strongest association of secPD-L1 with full-length PD-L1, but also with subsets of immunologic genes, such as in myeloid-derived suppressor cells. Moreover, the splice variant is also expressed in normal tissues and within normal peripheral blood cells it is preferentially expressed in activated myeloid cells. This is the first report of a form of secreted PD-L1 that homodimerizes and is functionally active. SecPD-L1 may function as a paracrine negative immune regulator within the tumor, since secPD-L1 does not require a cell-to-cell interaction to mediate its inhibitory effect. Electronic supplementary material The online version of this article (10.1007/s00262-018-2282-1) contains supplementary material, which is available to authorized users.

lsa(E) is a pleuromutilin, lincosamide, and streptogramin A (PLSA phenotype) resistance gene that was first described in S. aureus and was thought to have been transferred from Enterococcus sp. This study aimed to elucidate the prevalence of the lsa(E) gene among E. faecium isolates at a tertiary teaching hospital and to evaluate the transferability of the lsa(E) gene from E. faecium to S. aureus in vitro. A total of 96 E. faecium strains isolated from one hospital in Beijing in 2013 were analysed for quinupristin-dalfopristin (QDA) resistance genes, and multilocus sequence typing (MLST) was performed. The transferability of QDA resistance between ten E. faecium strains and four S. aureus strains was determined by filter mating. Genome sequencing of the transconjugant was performed. A total of 46 E. faecium isolates (46/96, 47.92%) tested positive for lsa(E), while two isolates (2/96, 2.08%) tested positive for lsa(A). Thirty-six lsa(E)-positive strains (36/46, 78.3%) belonged to ST78. Among 40 mating tests, lsa(E) was successfully transferred through one conjugation at a frequency of 1.125 × 10–7 transconjugants per donor. The QDA resistance of the transconjugant N7435-R3645 was expressed at a higher level (MIC = 16 mg/L) than that of the parent S. aureus strain (MIC = 0.38 mg/L). Next-generation sequencing (NGS) analysis of the transconjugant N7435-R3645 showed that the complete sequence of the lsa(E)-carrying plasmid pN7435-R3645 had a size of 92,396 bp and a G + C content of 33% (accession no. MT022086). The genetic map of pN7435-R3645 had high nucleotide similarity and shared the main open reading frame (ORF) features with two plasmids: E. faecium pMG1 (AB206333.1) and E. faecium LS170308 (CP025078.1). The rep gene of pN7435-R3645 showed 100% identity with that of pMG1, although it did not belong to the rep1-19 family but instead a unique rep family. Multiple antibiotic resistance genes, including lsa(E), aadE and lnu(B), erm(B), ant6-Ia, and lnu(B), were present on the plasmid. In conclusion, an lsa(E)-carrying plasmid that can be transferred by conjugation from E. faecium to S. aureus in vitro was identified. This multidrug resistance (MDR) pMG1-like plasmid may act as a vector in the dissemination of antimicrobial resistance among species.

BackgroundImmune checkpoint blockade (ICB) therapy has improved patient survival in a variety of cancers, but only a minority of cancer patients respond. Multiple studies have sought to identify general biomarkers of ICB response, but elucidating the molecular and cellular drivers of resistance for individual tumors remains challenging. We sought to determine whether a tumor with defined genetic background exhibits a stereotypic or heterogeneous response to ICB treatment.ResultsWe establish a unique mouse system that utilizes clonal tracing and mathematical modeling to monitor the growth of each cancer clone, as well as the bulk tumor, in response to ICB. We find that tumors derived from the same clonal populations showed heterogeneous ICB response and diverse response patterns. Primary response is associated with higher immune infiltration and leads to enrichment of pre-existing ICB-resistant cancer clones. We further identify several cancer cell-intrinsic gene expression signatures associated with ICB resistance, including increased interferon response genes and glucocorticoid response genes. These findings are supported by clinical data from ICB treatment cohorts.ConclusionsOur study demonstrates diverse response patterns from the same ancestor cancer cells in response to ICB. This suggests the value of monitoring clonal constitution and tumor microenvironment over time to optimize ICB response and to design new combination therapies. Furthermore, as ICB response may enrich for cancer cell-intrinsic resistance signatures, this can affect interpretations of tumor RNA-seq data for response-signature association studies.

Cancer treatment by immune checkpoint blockade (ICB) can bring long-lasting clinical benefits, but only a fraction of patients respond to treatment. To predict ICB response, we developed TIDE, a computational method to model two primary mechanisms of tumor immune evasion: the induction of T cell dysfunction in tumors with high infiltration of cytotoxic T lymphocytes (CTL) and the prevention of T cell infiltration in tumors with low CTL level. We identified signatures of T cell dysfunction from large tumor cohorts by testing how the expression of each gene in tumors interacts with the CTL infiltration level to influence patient survival. We also modeled factors that exclude T cell infiltration into tumors using expression signatures from immunosuppressive cells. Using this framework and pre-treatment RNA-Seq or NanoString tumor expression profiles, TIDE predicted the outcome of melanoma patients treated with first-line anti-PD1 or anti-CTLA4 more accurately than other biomarkers such as PD-L1 level and mutation load. TIDE also revealed new candidate ICB resistance regulators, such as SERPINB9, demonstrating utility for immunotherapy research.

In this paper, based on the nonlocal nonlinear Schrodinger equation, the evolution of complex-valued hyperbolic-sine-Gaussian beams (CVHSGBs) is investigated in nonlinear media with a spatial nonlocality. It is found that the evolution of CVHSGBs is variable depending on the parameters of complex-valued hyperbolic sine function. Choosing special parameters, the pattern of CVHSGBs can keep unchanged during propagation, and they propagate as solitons or breathers. Furthermore, for the general case, the CVHSGB evolutes periodically, and it recovers into its initial pattern at the end of each evolution period, namely it can be revivable periodically, which can be regarded as a generalized high-order breather. A series of analytical expressions are derived to describe the beam evolution, the intensity pattern, the beam spot size, the real beam curvature, etc. Some numerical simulations are also performed to demonstrate the typical evolution properties.

pH-responsive micelles with a biodegradable PLA core and a mixed PEG/PDPA shell were prepared by self-assembly of poly(ethylene glycol)-b-poly(lactic acid) (PEG-b-PLA) and poly(2-(diisopropylamino)ethyl methacrylate)-b-poly(lactic acid) (PDPA-b-PLA). The micellization status with different pH and the enzyme degradation behavior were characterized by 1H-NMR spectroscopy, dynamic light scattering measurement and zeta potential test. The pH turning point of PDPA block was determined to be in the range of 5.5−7.0. While the pH was above 7.0, the PDPA block collapsed onto the PLA core and could protect the PLA core from invasion of enzyme, as a result, the micelle exhibited a resistance to the enzymatic degradation.

The variation of roll restoring arm in waves is one of the critical factors for the occurrence of some dangerous stability failure modes, such as parametric rolling and pure loss of stability. This paper focuses on the accurate prediction of roll restoring arm in waves. Firstly, systematic partially restrained model tests of 75 experimental conditions at different heel angles, different forward speeds and different wave conditions are conducted by taking flared C11 containership under two drafts and ONR tumblehome hull as research objects. Secondly, detailed numerical simulation based on a three dimensional hybrid time domain panel methods are carried out, and the calculation results are benchmarked against model test data. Thirdly, the effects of different components of GZ are analyzed by comparing the numerical simulations with experimental results, in which the component induced by hydrostatic and Froude-Krylov forces (GZFK) are calculated by integrating pressure over the instantaneous wetted ship surface and the component induced by radiation and diffraction forces (GZRD) are calculated using the nonlinear body condition of the boundary-value problem. The investigations show that the conventional FK assumption in which GZRD is ignored can be employed for the calculation of roll restoring arm of zero speed in different wave directions, because GZRD is negligible at zero speed. Considering that there is no obvious improvement on the prediction accuracy in following waves with forward speed even though GZRD is taken into account, the FK assumption can also be used. The existence of GZRD is an indispensable part in head waves with forward speed, and the prediction accuracy of GZ can be drastically improved on the consideration of GZRD. Therefore, GZRD can be ignored for stability failure modes in following seas, while GZRD should be considered in head seas.

Immune checkpoint blockade (ICB) therapy revolutionized cancer treatment, but many patients with impaired MHC-I expression remain refractory. Here, we combined FACS-based genome-wide CRISPR screens with a data-mining approach to identify drugs that can upregulate MHC-I without inducing PD-L1. CRISPR screening identified TRAF3, a critical suppressor of the NF-κB pathway, as a negative regulator of MHC-I but not PD-L1. The Traf3-knockout (Traf3-KO) gene expression signature is associated with better survival in ICB-naive cancer patients and better ICB response. We then screened for drugs with similar transcriptional effects as this signature and identified SMAC mimetics. We experimentally validated that the SMAC mimetic birinapant upregulates MHC-I, sensitizes cancer cells to T-cell-dependent killing, and synergizes with ICB. Our findings provide preclinical rationale for treating tumors expressing low MHC-I expression with SMAC mimetics to enhance sensitivity to immunotherapy. The approach used in this study can be generalized to identify other drugs that enhance immunotherapy efficacy. Citation Format: Shengqing Gu, Wubing Zhang, Xiaoqing Wang, Peng Jiang, Nicole Traugh, Ziyi Li, Clifford Meyer, Blair Stewig, Yingtian Xie, Xia Bu, Michael Manos, Alba Font-Tello, Evisa Gjini, Ana Lako, Klothilda Lim, Jake Conway, Alok Tewari, Zexian Zeng, Avinash Das Sahu, Collin Tokheim, Jason L. Weirather, Jingxin Fu, Yi Zhang, Benjamin Kroger, Jin Hua Liang, Paloma Cejas, Gordon J. Freeman, Scott J. Rodig, Henry Long, Benjamin E. Gewurz, F. Stephen Hodi, Myles Brown, X. Shirley Liu. Therapeutically increasing MHC-I expression potentiates immune checkpoint blockade [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 65.

Aberrant energy status contributes to multiple metabolic diseases, including obesity, diabetes, and cancer, but the underlying mechanism remains elusive. Here, we report that ketogenic-diet-induced changes in energy status enhance the efficacy of anti-CTLA-4 immunotherapy by decreasing PD-L1 protein levels and increasing expression of type-I interferon (IFN) and antigen presentation genes. Mechanistically, energy deprivation activates AMP-activated protein kinase (AMPK), which in turn, phosphorylates PD-L1 on Ser283, thereby disrupting its interaction with CMTM4 and subsequently triggering PD-L1 degradation. In addition, AMPK phosphorylates EZH2, which disrupts PRC2 function, leading to enhanced IFNs and antigen presentation gene expression. Through these mechanisms, AMPK agonists or ketogenic diets enhance the efficacy of anti-CTLA-4 immunotherapy and improve the overall survival rate in syngeneic mouse tumor models. Our findings reveal a pivotal role for AMPK in regulating the immune response to immune-checkpoint blockade and advocate for combining ketogenic diets or AMPK agonists with anti-CTLA4 immunotherapy to combat cancer.

There is one project related to high resolution aeronautic camera, which is used in the Military aircraft. In the project, the task requires that the camera's field angle is ±20 degrees. Besides, we there need a window glass for the camera, and the distance between the window-glass and the platform is limited not more than 30mm. That is to say, the aerial camera is required to have a large field of view and small optical window order. To solve this problem, we designed a four-section-type optical window. This paper mainly describes the possible kinds of joining of the glasses used in this four-section-type optical window. There are three kinds of joining mentioned in this paper, such as mechanical connection, joining with adhesive and welding connection. Every kind has its own advantages and disadvantages. After the comprehensive consideration, the paper will use the method of mechanical connection. And this paper will give some guiding significance for the subsequent progress of the project.

In this paper, the evolution dynamics of astigmatic four-petal Gaussian beams in isotropic nonlocal nonlinear media is investigated. A series of analytical expressions are derived to describe the beam evolution, the beam width, and the critical power. The anomalous transverse transformation is presented, i.e., an astigmatic four-petal Gaussian beam is compressed in one transverse direction and it is broadened in the other transverse direction. These evolution characteristics of astigmatic four-petal Gaussian beams are illustrated through some numerical simulations. Keywords: Isotropic nonlocal media, Astigmatic four-petal Gaussian beam, Anomalous transverse transformation

Immunotherapy using checkpoint-blocking antibodies against PD-1 has produced impressive results in a wide range of cancers. However, the response remains heterogeneous among patients. We used noninvasive immuno-positron emission tomography (PET), using (89)Zr-labeled PEGylated single-domain antibody fragments (nanobodies or VHHs), to explore the dynamics and distribution of intratumoral CD8(+) T cells and CD11b(+) myeloid cells in response to anti–PD-1 treatment in the MC38 colorectal mouse adenocarcinoma model. Responding and nonresponding tumors showed consistent differences in the distribution of CD8(+) and CD11b(+) cells. Anti–PD-1 treatment mobilized CD8(+) T cells from the tumor periphery to a more central location. Only those tumors fully infiltrated by CD8(+) T cells went on to complete resolution. All tumors contained CD11b(+) myeloid cells from the outset of treatment, with later recruitment of additional CD11b(+) cells. As tumors grew, the distribution of intratumoral CD11b(+) cells became more heterogeneous. Shrinkage of tumors in responders correlated with an increase in the CD11b(+) population in the center of the tumors. The changes in distribution of CD8(+) and CD11b(+) cells, as assessed by PET, served as biomarkers to gauge the efficacy of anti–PD-1 treatment. Single-cell RNA sequencing of RNA from intratumoral CD45(+) cells showed that CD11b(+) cells in responders and nonresponders were markedly different. The responders exhibited a dominant population of macrophages with an M1-like signature, while the CD45(+) population in the nonresponders displayed an M2-like transcriptional signature. Thus, by using immuno-PET and single-cell RNA sequencing, we show that anti–PD-1 treatment not only affects interactions of CD8(+) T cells with the tumor but also impacts the intratumoral myeloid compartment.

The aim of the present study was to investigate the effect of Scutellaria baicalensis stem-leaf total flavonoid (SSTF) on the dopaminergic neurons in the substantia nigra in a mouse model of Parkinson's disease (PD). The mouse model was established by intravenous injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). SSTF (5 mg/kg) was administered to the mice before or after MPTP injection, and the effects of SSTF on the behavior of the mice and the dopaminergic neurons in the substantia nigra were assessed. In addition, the level of serum malondialdehyde (MDA) was measured. Following injection of MPTP, the number of dopaminergic neurons in the substantia nigra was decreased and the neurons appeared atrophic. In addition, the level of serum MDA in the MPTP mice increased. The mean behavioral scores and the number of dopaminergic neurons in the SSTF treatment groups were significantly higher than in the MPTP group (P

Sparse signal recovery is attractive in compressed sensing (CS). Based on the smoothed ?0 norm (SL0) algorithm, we have developed an error-tolerant regularized SL0 (ReSL0) algorithm, which has the same computational advantages as the SL0 algorithm while having better immunity against inaccuracy caused by noise or model mismatch. The performance of the ReSL0 is evaluated with simulated data. In addition, we have extended the ReSL0 to the matrix form (MReSL0), which is more suitable for dealing with matrix form signals and also has good resilience against inaccuracy. Finally we apply the ReSL0 and MReSL0 to joint CS-based radar imaging and phase error correction. Experimental results from both simulated and real data demonstrate that the proposed algorithms provide remarkable performance improvements in inaccurate scenarios (such as noisy data and mismatched settings) compared with the SL0 algorithm. HighlightsAn error-tolerant regularized SL0 algorithm is proposed to recover sparse signals.The matrix form regularized SL0 algorithm is derived to recover matrix form signals.The above algorithms are applied to joint CS-based radar imaging and phase error correction.

Blockade of PD-1/PD-L1 is in effective anti-tumor immunotherapy. To further elucidate the mechanism by which PD-1/PD-L1 engagement causes T cell dysfunction, we investigated intracellular signaling triggered by PD-1/PD-L1 ligation in T cells. We describe a novel antibody that detects PD-1 signaling. This antibody detects phosphorylation of the immunotyrosine switch motif (ITSM) in the intracellular tail of both human and mouse PD-1 (phospho-PD-1). We first examined the expression of PD-1 and phosphorylated PD-1 in T cells from human PBMC. We found that without CD3 and CD28 mAb stimulation, PD-1 and phospho-PD-1 had minimal expression on CD3+CD4+ and CD3+CD8+ cell. Upon stimulation with CD3 and CD28 mAb, we observed increased expression of PD-1 and phospho PD-1 in both CD3+CD4+ and CD3+CD8+ cells. Furthermore, upon CD3 and CD28 mAb stimulation and PD-L1 recombinant protein treatment, the level of phospho PD-1 increased in CD3+CD4+ and CD3+CD8+ cells. Utilizing a co-culture system of Jurkat-hPD-1 and 300-hPD-L1, we mimicked the cell-cell interaction through molecular ligation of PD-1 and PD-L1. Upon stimulation with CD3 and CD28 mAb in this co-culture system, we detected increased phospho PD-1 signal and phospho SHP-2 signal by Western blot. When CD3 and CD28 mAb stimulated Jurkat-hPD-1 cells were treated with recombinant PD-L1 protein, we observed decreased IL-2 secretion which indicates PD-1/PD-L1 ligation suppresses T cell activation and corresponds with phospho PD-1 expression. This study examines the intrinsic intracellular signals upon PD-1/PD-L1 engagement, and the results may promote a better understanding of the mechanism of T cell dysfunction by PD-1/PD-L1 ligation.

Therapeutic cancer vaccines aim to treat pre-existing cancer by boosting the patient's own immune system, which is an attractive strategy for cancer treatment. The cancer vaccines have mainly been designed to elicit antitumor T-cell immune responses that recognize and eradicate cancer. The advantages of cancer immunotherapy with cancer vaccines include a) high specificity of tumor antigen, b) minimal vaccine-related adverse events, and c) long-lasting immunity boosted by cancer vaccine which is important to control tumor relapse. In this chapter, we discuss identification of tumor antigens in breast cancer (e.g., cancer-testis antigens, neoantigens, HER2/neu, MUC1), the vaccine delivery systems utilized in breast cancer treatment (e.g., peptide vaccines, dendritic cell-based vaccines, and whole tumor cell-based vaccines), as well as clinical trials with therapeutic breast cancer vaccines. Moreover, new-generation clinical trials of breast cancer vaccines will aim at employing personalized vaccines designed to harness robust immune response to a custom-made neoantigen in the patient with breast cancer. Combination of vaccination and other forms of cancer therapy such as chemotherapy, radiotherapy, targeted therapy with monoclonal antibody, or immune checkpoint blockade will be required to achieve potent and durable antitumor clinical benefits.

Cancer immunotherapy is emerging as the most promising novel strategy for cancer treatment. Cancer immunotherapy is broadly categorized into three forms: immune checkpoint modulation, adoptive cell transfer, and cancer vaccine. Immune checkpoint blockade is demonstrated as the most clinically effective treatment with low immune-related adverse events (irAE). Blockade of PD-1/PD-L1 and CTLA-4 has achieved remarkable success in treating various types of tumors, which sparks great interests in this therapeutic strategy and expands the role of immune checkpoint blockade in treating tumors including breast cancer. Based on the notable results obtained from clinical trials, the United States' Food and Drug Administration (FDA) has approved multiple CTLA-4 monoclonal antibodies as well as the PD-1/PD-L1 monoclonal antibodies for treatment of different types of tumors. The theories of immunoediting, T-cell exhaustions, and co-stimulatory/co-inhibitory pathways are immunological foundations for immune checkpoint blockade therapy. Breast cancers such as triple negative breast cancer and HER-2 negative breast cancer respond to immune checkpoint blockade therapy due to their high immunogenicity. PD-1/PD-L1 blockade has just received FDA approval as a standard cancer therapy for solid tumors such as breast cancer. Development of immune checkpoint blockade focuses on two directions: one is to identify proper biomarkers of immune checkpoint blockade in breast cancer, and the other is to combine therapies with PD-1/PD-L1 blockade antibodies to achieve optimal clinical outcomes.

The study has investigated the effect of isoflavone attenuates the caspase-1 and caspase-3 level in cell model of Parkinsonism. The subjects were PC12 cells. They were randomly divided into six groups: control, MPP+(250 μmol/L), isoflavone (10 μM), isoflavone (10 μM) + MPP+(250 μmol/L), Z-YVAD-CHO (10 nM) + MPP+group, and Z-DEVD-CHO (10 nM) + MPP+group. Cell viability was measured by MTT methods; the content of tyrosine hydroxylase was measured by immunocytochemistry method of avidinbiotin peroxidase complex; apoptosis ratio was measured by flow cytometry. The results showed that cell viability in the MPP+group was lower than in all other five groups. There was no difference in cell viability between isoflavone + MPP+and control group. Optical density of TH positive cells in isoflavone group was higher than in control, isoflavone + MPP+, and MPP+only groups. The apoptosis ratio in the isoflavone + MPP+group and control group and the Z-YVAD-CHO + MPP+and Z-DEVD-CHO + MPP+group was similar, which was lower than in the MPP+group. The lowest apoptosis ratio was found in the isoflavone only group.

Treatments that target immune checkpoints, such as the one mediated by programmed cell death protein 1 (PD-1) and its ligand PD-L1, have been approved for treating human cancers with durable clinical benefit. However, many patients with cancer fail to respond to compounds that target the PD-1 and PD-L1 interaction, and the underlying mechanism(s) is not well understood. Recent studies revealed that response to PD-1-PD-L1 blockade might correlate with PD-L1 expression levels in tumour cells. Hence, it is important to understand the mechanistic pathways that control PD-L1 protein expression and stability, which can offer a molecular basis to improve the clinical response rate and efficacy of PD-1-PD-L1 blockade in patients with cancer. Here we show that PD-L1 protein abundance is regulated by cyclin D-CDK4 and the cullin 3-SPOP E3 ligase via proteasome-mediated degradation. Inhibition of CDK4 and CDK6 (hereafter CDK4/6) in vivo increases PD-L1 protein levels by impeding cyclin D-CDK4-mediated phosphorylation of speckle-type POZ protein (SPOP) and thereby promoting SPOP degradation by the anaphase-promoting complex activator FZR1. Loss-of-function mutations in SPOP compromise ubiquitination-mediated PD-L1 degradation, leading to increased PD-L1 levels and reduced numbers of tumour-infiltrating lymphocytes in mouse tumours and in primary human prostate cancer specimens. Notably, combining CDK4/6 inhibitor treatment with anti-PD-1 immunotherapy enhances tumour regression and markedly improves overall survival rates in mouse tumour models. Our study uncovers a novel molecular mechanism for regulating PD-L1 protein stability by a cell cycle kinase and reveals the potential for using combination treatment with CDK4/6 inhibitors and PD-1-PD-L1 immune checkpoint blockade to enhance therapeutic efficacy for human cancers.

Cancer immunotherapy is emerging as the most promising novel strategy for cancer treatment. Cancer immunotherapy is broadly categorized into three forms: immune checkpoint modulation, adoptive cell transfer, and cancer vaccine. Immune checkpoint blockade is demonstrated as the most clinically effective treatment with low immune-related adverse events (irAE). Blockade of PD-1/PD-L1 and CTLA-4 has achieved remarkable success in treating various types of tumors, which sparks great interests in this therapeutic strategy and expands the role of immune checkpoint blockade in treating tumors including breast cancer. Based on the notable results obtained from clinical trials, the United States’ Food and Drug Administration (FDA) has approved multiple CTLA-4 monoclonal antibodies as well as the PD-1/PD-L1 monoclonal antibodies for treatment of different types of tumors. The theories of immunoediting, T-cell exhaustions, and co-stimulatory/co-inhibitory pathways are immunological foundations for immune checkpoint blockade therapy. Breast cancers such as triple negative breast cancer and HER-2 negative breast cancer respond to immune checkpoint blockade therapy due to their high immunogenicity. PD-1/PD-L1 blockade has just received FDA approval as a standard cancer therapy for solid tumors such as breast cancer. Development of immune checkpoint blockade focuses on two directions: one is to identify proper biomarkers of immune checkpoint blockade in breast cancer, and the other is to combine therapies with PD-1/PD-L1 blockade antibodies to achieve optimal clinical outcomes.

In generally, there are multiple factories. According to mining areas, it supplies different quality iron ore concentrate. And according to the unique requirements of owners, different grades concentrate cannot be confused. Therefore, the grade of slurry pipeline transportation is needed to be used. When in the study of grade transmission, we must focus on Hydraulic characteristics of solid-liquid two-phase for different quality of iron ore concentrate. Slurry concentration and Critical velocity of flow is especially important. Research shows that the deposition rate of laminar or turbulent transition velocity is higher than the occasion when the slurry has concentration, so the minimum operating speed is controlled by sedimentation; the concentration lower limit is 62%. If the concentration is over 68%, the yield stress of slurry will increase, and the pipeline pressure will drop, which is very sensitive to slight changes in concentration. Therefore, the solid weight concentration range is 62% to 68%,as the selection range of design.

In view of the characteristics of the non-stationary and multi-component AM-FM signals of vibration signals in the rolling element bearing, the generalized demodulation time-frequency analysis method is used for its fault diagnosis, overcoming the problem that the maximal overlap discrete wavelet packet transform (MODWPT) has no adaptability. First of all, the original vibration signal is took preprocessing by generalized Fourier; Then, using MODWPT to decompose signals after pretreatment and obtaining weights; Once again, the weights are carried out the inverse generalized Fourier transform to get the weights of the original signal; Finally, reconstructing principal component of the original signal to get the Hilbert instantaneous energy spectrum, roller bearing fault diagnoses based on the Hilbert instantaneous energy spectrum. The experimental results show that the method can effectively diagnose rolling bearing fault.