Publications by authors named "Guoan Zhang"

89 Publications

Fascin inhibitor increases intratumoral dendritic cell activation and anti-cancer immunity.

Cell Rep 2021 Apr;35(1):108948

Department of Physiology and Biophysics, Weill Cornell Medical College of Cornell University, New York, NY 10065, USA; Sandra and Edward Meyer Cancer Center, Weill Cornell Medicine of Cornell University, New York, NY 10065, USA. Electronic address:

Fascin protein is the main actin-bundling protein in filopodia and invadopodia, which are critical for tumor cell migration, invasion, and metastasis. Small-molecule fascin inhibitors block tumor invasion and metastasis and increase the overall survival of tumor-bearing mice. Here, we report a finding that fascin blockade additionally reinvigorates anti-tumor immune response in syngeneic mouse models of various cancers. Fascin protein levels are increased in conventional dendritic cells (cDCs) in the tumor microenvironment. Mechanistically, fascin inhibitor NP-G2-044 increases the number of intratumoral-activated cDCs and enhances the antigen uptake by cDCs. Furthermore, together with PD-1 blocking antibody, NP-G2-044 markedly increases the number of activated CD8 T cells in the otherwise anti-PD-1 refractory tumors. Reduction of fascin levels in cDCs, but not fascin gene knockout in tumor cells, mimics the anti-tumor immune effect of NP-G2-044. These data demonstrate that fascin inhibitor NP-G2-044 simultaneously limits tumor metastasis and reinvigorates anti-tumor immune responses.
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http://dx.doi.org/10.1016/j.celrep.2021.108948DOI Listing
April 2021

Combining the advantages of prokaryotic expression and T7 phage display systems to obtain antigens for antibody preparation.

Protein Expr Purif 2020 Dec 11;184:105808. Epub 2020 Dec 11.

Cangzhou Nanobody Technology Innovation Center, Cangzhou Medical College, Cangzhou, 061001, China. Electronic address:

The gene encoding the phage major capsid protein 10A was cloned into the prokaryotic expression vector pET24a, and a 6XHis-tag was fused to the 3'-end of the 10A gene to verify complete expression. The recombinant plasmid was transformed into Escherichia coli (E. coli) BL21 (DE3) cells, and 10A expression was induced by IPTG. SDS-PAGE and Western blot were used to confirm the target protein expression. The T7Select10-3b vector was added to the cultured bacteria expressing 10A at a multiplicity of infection (MOI) ranging from 0.01 to 0.1, and complete lysis of the bacteria was monitored by absorbance changes in the medium. The recombinant phage (reP) was harvested by PEG/NaCl sedimentation and resuspended in PBS. ELISA was performed to verify the presence of the 6XHis-tag on the surface of reP. The 10A-fusion expression vectors (pET10A-flag, pET10A-egfp, and pET10A-pct) were constructed, and fusion proteins were expressed and detected by the same method. The corresponding rePs (reP-Flag, reP-EGFP, and reP-PCT) were prepared by T7Select10-3b infection. After the expression of the peptides/proteins on the reP surfaces was confirmed, reP-Flag and reP-PCT were used to immunize mice to prepare anti-Flag and anti-PCT antibodies. The results showed that rePs prepared using the 10A-fusion vector and T7Select10-3b can be used as antigens to immunize mice and prepare antibodies. This method may be able to meet the rapid antigen preparation requirements for antibody production. Notably, the recombinant phage (reP) described in this study was obtained by the sedimentation method from T7Select10-3b-infected E. coli BL21 (DE3) cells carrying the major capsid protein 10A expression vector or 10A-fusion protein vector.
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http://dx.doi.org/10.1016/j.pep.2020.105808DOI Listing
December 2020

Proteasome inhibitors attenuates mitoxantrone-triggered immunogenic cell death in prostate cancer cells.

Med Oncol 2020 Nov 19;37(12):116. Epub 2020 Nov 19.

Institute of Precision Medicine, Jining Medical University, Jining, 272067, China.

Both mitoxantrone (MTX) and proteasome inhibitors efficiently trigger immunogenic cell death (ICD) in cancer cells. However, whether the combination of MTX and proteasome inhibitors can synergistically enhance ICD remains unknown. In this study, we showed that the proteasome inhibitors bortezomib (BZM) and carbobenzoxy-L-leucyl-L-leucyl-L-leucinal (MG132) impaired MTX-induced ICD in prostate cancer cells, as measured using ICD biomarkers and dendritic cell activation in vitro. Mice vaccinated with RM-1 mouse prostate cancer cell line treated with BZM or MG132 in combination with MTX showed enhanced tumor growth, and shortened tumor-free, and worse overall survival compared with those treated with MTX alone. In conclusion, we demonstrated that proteasome inhibitors (BZM or MG132) attenuated MTX-induced ICD, suggesting that proteasome activation was required for MTX-induced ICD.
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http://dx.doi.org/10.1007/s12032-020-01445-yDOI Listing
November 2020

Pharmaceutical inhibition of AXL suppresses tumor growth and invasion of esophageal squamous cell carcinoma.

Exp Ther Med 2020 Nov 2;20(5):41. Epub 2020 Sep 2.

Information Technology Centre, Jining Medical University, Jining, Shandong 272067, P.R. China.

Esophageal squamous cell carcinoma (ESCC) is a common type of cancer in a number of regions of the world, including East Asia, South Africa and Iran. It is often associated with poor prognosis rates. Tyrosine-protein kinase receptor UFO (AXL) is overexpressed in a subset of ESCC tumors, therefore the present study aimed to determine the effect of R428, a selective inhibitor of AXL, on ESCC tumor cells. TE1 and KYSE150 cell lines were used as models to investigate the effects of R428 treatment. The proliferative rate of the tumor cells was analyzed using MTT and colony formation assays. In addition, cell migration and invasion rates were analyzed using wound healing and Matrigel assays, respectively. The expression levels of matrix metalloproteinase (MMP)2 and MMP9, and the activation of protein kinase B (AKT), extracellular signal-regulated kinase (ERK) and AXL signaling were analyzed using gelatin zymography and western blotting. The results revealed that R428 inhibited the proliferative and invasive abilities of both cell lines. Furthermore, AXL, AKT and ERK signaling were all decreased in response to R428 treatment, alongside the expression levels of MMP2 and MMP9. In conclusion, the results of the present study suggested that R428 treatment may suppress ESCC tumor cell proliferation and invasion, representing a potential therapeutic target for ESCC.
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http://dx.doi.org/10.3892/etm.2020.9169DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7480165PMC
November 2020

PET, image-guided HDAC inhibition of pediatric diffuse midline glioma improves survival in murine models.

Sci Adv 2020 Jul 24;6(30):eabb4105. Epub 2020 Jul 24.

Department of Neurological Surgery, Weill Cornell Medicine, New York, NY 10065, USA.

Efforts at altering the dismal prognosis of pediatric midline gliomas focus on direct delivery strategies like convection-enhanced delivery (CED), where a cannula is implanted into tumor. Successful CED treatments require confirmation of tumor coverage, dosimetry, and longitudinal in vivo pharmacokinetic monitoring. These properties would be best determined clinically with image-guided dosimetry using theranostic agents. In this study, we combine CED with novel, molecular-grade positron emission tomography (PET) imaging and show how PETobinostat, a novel PET-imageable HDAC inhibitor, is effective against DIPG models. PET data reveal that CED has significant mouse-to-mouse variability; imaging is used to modulate CED infusions to maximize tumor saturation. The use of PET-guided CED results in survival prolongation in mouse models; imaging shows the need of CED to achieve high brain concentrations. This work demonstrates how personalized image-guided drug delivery may be useful in potentiating CED-based treatment algorithms and supports a foundation for clinical translation of PETobinostat.
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http://dx.doi.org/10.1126/sciadv.abb4105DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7439439PMC
July 2020

SDN-Based Survivability Analysis for V2I Communications.

Sensors (Basel) 2020 Aug 19;20(17). Epub 2020 Aug 19.

School of Information Science and Technology, Nantong University, Nantong 226019, China.

In vehicle-to-infrastructure (V2I) communications, various failures in the dynamic movement pose serious link interruptions. To study the continuous service quality for the V2I network when this issue happens, this paper proposes a survivability analysis and establishes the communication architecture for software-defined network (SDN)-based V2I communications. With the controllable advantages of SDN centralized management, the multi-path transmission control protocol is used to seamlessly switch the transmission information between the V2I links of each vehicle node. Specifically, according to the analysis of specific fault types for V2I links, the definitions of SDN-based V2I survivability is provided to establish the corresponding survivability mode. To further verify the survivability model, a full-state search is adopted by means of probability model checker PRISM. In addition, multi-directional probability and expected reward evaluation analyses are carried out from the point of view of time. The simulation results show that, with the failure of multiple V2I links, the network quality of service (QoS) correspondingly declines, but the network still survives, due to the multi-path transmission control protocol (MPTCP) action. Moreover, with a high fault repair rate, the service performance and survivability of the network is improved rapidly.
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http://dx.doi.org/10.3390/s20174678DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7506569PMC
August 2020

NRH salvage and conversion to NAD requires NRH kinase activity by adenosine kinase.

Nat Metab 2020 04 21;2(4):364-379. Epub 2020 Apr 21.

Department of Pharmacology, Weill Cornell Medical College, New York, NY, USA.

Dihydronicotinamide riboside (NRH) has been suggested to act as a precursor for the synthesis of NAD, but the biochemical pathway converting it has been unknown. Here, we show that NRH can be converted into NAD via a salvage pathway in which adenosine kinase (ADK, also known as AK) acts as an NRH kinase. Using isotope-labelling approaches, we demonstrate that NRH is fully incorporated into NAD, with NMNH acting as an intermediate. We further show that AK is enriched in fractions from cell lysates with NRH kinase activity, and that AK can convert NRH into NAD. In cultured cells and mouse liver, pharmacological or genetic inhibition of AK blocks formation of reduced nicotinamide mononucleotide (NMNH) and inhibits NRH-stimulated NAD biosynthesis. Finally, we confirm the presence of endogenous NRH in the liver with metabolomics. Our findings establish NRH as a natural precursor of NAD and reveal a new route for NAD biosynthesis via an NRH salvage pathway involving AK.
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http://dx.doi.org/10.1038/s42255-020-0194-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7384296PMC
April 2020

Neuronal Inactivity Co-opts LTP Machinery to Drive Potassium Channel Splicing and Homeostatic Spike Widening.

Cell 2020 06 2;181(7):1547-1565.e15. Epub 2020 Jun 2.

Department of Neuroscience and Physiology, Neuroscience Institute, NYU Grossman Medical Center, New York, NY 10016, USA; Center for Neural Science, New York University, New York, NY 10003, USA. Electronic address:

Homeostasis of neural firing properties is important in stabilizing neuronal circuitry, but how such plasticity might depend on alternative splicing is not known. Here we report that chronic inactivity homeostatically increases action potential duration by changing alternative splicing of BK channels; this requires nuclear export of the splicing factor Nova-2. Inactivity and Nova-2 relocation were connected by a novel synapto-nuclear signaling pathway that surprisingly invoked mechanisms akin to Hebbian plasticity: Ca-permeable AMPA receptor upregulation, L-type Ca channel activation, enhanced spine Ca transients, nuclear translocation of a CaM shuttle, and nuclear CaMKIV activation. These findings not only uncover commonalities between homeostatic and Hebbian plasticity but also connect homeostatic regulation of synaptic transmission and neuronal excitability. The signaling cascade provides a full-loop mechanism for a classic autoregulatory feedback loop proposed ∼25 years ago. Each element of the loop has been implicated previously in neuropsychiatric disease.
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http://dx.doi.org/10.1016/j.cell.2020.05.013DOI Listing
June 2020

Energy-Optimal Latency-Constrained Application Offloading in Mobile-Edge Computing.

Sensors (Basel) 2020 May 28;20(11). Epub 2020 May 28.

School of information science and technology, Nantong University, Nantong 226019, China.

Mobile-edge computation offloading (MECO) is a promising emerging technology for battery savings in mobile devices (MD) and/or in latency reduction in the execution of applications by (either total or partial) offloading highly demanding applications from MDs to nearby servers such as base stations. In this paper, we provide an offloading strategy for the joint optimization of the communication and computational resources by considering the blue trade-off between energy consumption and latency. The strategy is formulated as the solution to an optimization problem that minimizes the total energy consumption while satisfying the execution delay limit (or deadline). In the solution, the optimal transmission power and rate and the optimal fraction of the task to be offloaded are analytically derived to meet the optimization objective. We further establish the conditions under which the binary decisions (full-offloading and no offloading) are optimal. We also explore how such system parameters as the latency constraint, task complexity, and local computing power affect the offloading strategy. Finally, the simulation results demonstrate the behavior of the proposed strategy and verify its energy efficiency.
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http://dx.doi.org/10.3390/s20113064DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7309048PMC
May 2020

Pkm2 Regulates Cardiomyocyte Cell Cycle and Promotes Cardiac Regeneration.

Circulation 2020 04 11;141(15):1249-1265. Epub 2020 Feb 11.

Cardiovascular Research Center (A.M, N.S., A.A.K., I.M., T.M. K.B., M.T.K.S., E.C., Y.S., J.G.O., P.L, A.G.-S., C.K., M.M., L.Z.), Icahn School of Medicine at Mount Sinai, New York.

Background: The adult mammalian heart has limited regenerative capacity, mostly attributable to postnatal cardiomyocyte cell cycle arrest. In the last 2 decades, numerous studies have explored cardiomyocyte cell cycle regulatory mechanisms to enhance myocardial regeneration after myocardial infarction. Pkm2 (Pyruvate kinase muscle isoenzyme 2) is an isoenzyme of the glycolytic enzyme pyruvate kinase. The role of Pkm2 in cardiomyocyte proliferation, heart development, and cardiac regeneration is unknown.

Methods: We investigated the effect of Pkm2 in cardiomyocytes through models of loss (cardiomyocyte-specific Pkm2 deletion during cardiac development) or gain using cardiomyocyte-specific Pkm2 modified mRNA to evaluate Pkm2 function and regenerative affects after acute or chronic myocardial infarction in mice.

Results: Here, we identify Pkm2 as an important regulator of the cardiomyocyte cell cycle. We show that Pkm2 is expressed in cardiomyocytes during development and immediately after birth but not during adulthood. Loss of function studies show that cardiomyocyte-specific Pkm2 deletion during cardiac development resulted in significantly reduced cardiomyocyte cell cycle, cardiomyocyte numbers, and myocardial size. In addition, using cardiomyocyte-specific Pkm2 modified RNA, our novel cardiomyocyte-targeted strategy, after acute or chronic myocardial infarction, resulted in increased cardiomyocyte cell division, enhanced cardiac function, and improved long-term survival. We mechanistically show that Pkm2 regulates the cardiomyocyte cell cycle and reduces oxidative stress damage through anabolic pathways and β-catenin.

Conclusions: We demonstrate that Pkm2 is an important intrinsic regulator of the cardiomyocyte cell cycle and oxidative stress, and highlight its therapeutic potential using cardiomyocyte-specific Pkm2 modified RNA as a gene delivery platform.
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http://dx.doi.org/10.1161/CIRCULATIONAHA.119.043067DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7241614PMC
April 2020

Evaluation of allelic alterations in short tandem repeats in papillary thyroid cancer.

Mol Genet Genomic Med 2020 04 11;8(4):e1164. Epub 2020 Feb 11.

Institute of Forensic Medicine and Laboratory Medicine, Jining Medical University, Forensic Science Center of Jining Medical University, Jining, Shandong, PR China.

Background: Malignant tissue samples may be the only source of biological material for forensic investigations, including individual identification or paternity testing; however, such samples may lead to uncertainties due to frequent genomic aberrations associated with tumors, including alterations of the short tandem repeat (STR) loci used for forensic casework.

Methods: Short tandem repeat loci routinely used in forensic analysis (n = 23) were analyzed in 68 surgically removed papillary thyroid cancer specimens. Tumor cells and normal stromal cells were separated by laser capture microdissection.

Results: Four kinds of changes were detected between normal and tumor tissues: partial loss of heterozygosity (pLOH), complete loss of heterozygosity, an additional allele, and a new allele not found in normal tissue. These changes were distributed across 20 of the tested STRs, with no mutations in VWA, D16S539, or Penta D. The most frequently affected locus was D2S1338, and the most frequent type of alteration was pLOH. Samples from patients aged 40-59 years exhibited the highest frequencies of STR variation.

Conclusion: Our results suggest that great care should be taken in the evaluation of DNA typing results obtained from malignant tissues, particularly when no normal tissue reference samples are available.
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http://dx.doi.org/10.1002/mgg3.1164DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7196467PMC
April 2020

Population genetic data from 23 autosomal STR loci of Huaxia Platinum system in the Jining Han population.

Mol Genet Genomic Med 2020 04 29;8(4):e1142. Epub 2020 Jan 29.

Institute of Forensic Medicine and Laboratory Medicine, Jining Medical University, Jining, Shandong, PR China.

Background: Genetic polymorphisms at 23 short tandem repeat (STR) loci were investigated in 1,215 Jining Han individuals from Jining city, Shandong province, eastern China.

Methods: We used population genetic data of 23 autosomal STR loci included in the Huaxia Platinum system to evaluate 1,215 unrelated Chinese Han individuals in the Jining Han population. Allele frequencies and forensic parameters of the STR loci were determined and genetic relationships among the Jining Han and other Chinese populations were evaluated.

Results: In total, we observed 321 alleles, with frequencies ranging from 0.00041 to 0.52222. The combined discrimination power and probability of excluding paternity were 0.99999999999999999999999999919 and 0.99999999962, respectively. No deviations from HWE were observed at any loci. Population comparisons showed that the Xinjiang groups (Uyghur and Kazakh) and the Mongolian and Tibetan groups were isolated, while the Jining Han population clustered together with other populations, except the Guizhou Han population.

Conclusion: This study demonstrated that 23 autosomal STR loci included in the Huaxia Platinum system are highly polymorphic and suitable for personal forensic identification and paternity testing in this population.
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http://dx.doi.org/10.1002/mgg3.1142DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7196486PMC
April 2020

CSNK2A1 Promotes Gastric Cancer Invasion Through the PI3K-Akt-mTOR Signaling Pathway.

Cancer Manag Res 2019 2;11:10135-10143. Epub 2019 Dec 2.

Department of Oncology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong 250021, People's Republic of China.

Objective: Casein kinase 2 a1 (CSNK2A1) has been shown to be involved in tumorigenesis by enhancing several oncogenic signaling pathways in various cancers. However, the function and mechanism of CSNK2A1 in gastric cancer remain unclear, and this study aimed to elucidate the role of CSNK2A1 in gastric cancer.

Methods: CSNK2A1 expression was assessed by Western blot and qPCR in four gastric cancer (GC) cell lines and one normal gastric epithelial cell line. Stable cancer cell lines with CSNK2A1 gene overexpression or knockdown were established to investigate the function and mechanism of CSNK2A1 in GC cells.

Results: CSNK2A1 expression was higher in GC cells than in normal gastric epithelial cells. Stable overexpression of CSNK2A1 in SNU216 cells significantly increased cellular proliferation, invasion, and migration. Silencing CSNK2A1 expression in SGC-790 cells effectively inhibited its oncogenic function. We further verified that epithelial-mesenchymal transition (EMT) was affected by CSNK2A1 and that CSNK2A1 promotes GC cell invasion through the PI3K-Akt-mTOR signaling pathway.

Conclusion: Our findings suggested that CSNK2A1 plays important oncogenic roles in GC invasion via EMT and the PI3K-Akt-mTOR signaling pathway and that CSNK2A1 may serve as a novel prognostic and/or therapeutic target in GC.
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http://dx.doi.org/10.2147/CMAR.S222620DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6897054PMC
December 2019

Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) for Quantitative Proteomics.

Adv Exp Med Biol 2019 ;1140:531-539

Kimmel Center for Biology and Medicine at the Skirball Institute and Department of Cell Biology, New York University School of Medicine, New York, NY, USA.

Stable isotope labeling by amino acids in cell culture (SILAC) is a powerful approach for high-throughput quantitative proteomics. SILAC allows highly accurate protein quantitation through metabolic encoding of whole cell proteomes using stable isotope labeled amino acids. Since its introduction in 2002, SILAC has become increasingly popular. In this chapter we review the methodology and application of SILAC, with an emphasis on three research areas: dynamics of posttranslational modifications, protein-protein interactions, and protein turnover.
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http://dx.doi.org/10.1007/978-3-030-15950-4_31DOI Listing
September 2019

Inhibition of 3-phosphoglycerate dehydrogenase (PHGDH) by indole amides abrogates de novo serine synthesis in cancer cells.

Bioorg Med Chem Lett 2019 09 6;29(17):2503-2510. Epub 2019 Jul 6.

Meyer Cancer Center, Weill Cornell Medical College, New York, NY 10065, United States; Department of Medicine, Weill Cornell Medical College, New York, NY 10065, United States. Electronic address:

Cancer cells reprogram their metabolism to support growth and to mitigate cellular stressors. The serine synthesis pathway has been identified as a metabolic pathway frequently altered in cancers and there has been considerable interest in developing pharmacological agents to target this pathway. Here, we report a series of indole amides that inhibit human 3-phosphoglycerate dehydrogenase (PHGDH), the enzyme that catalyzes the first committed step of the serine synthesis pathway. Using X-ray crystallography, we show that the indole amides bind the NAD pocket of PHGDH. Through structure-based optimization we were able to develop compounds with low nanomolar affinities for PHGDH in an enzymatic IC assay. In cellular assays, the most potent compounds inhibited de novo serine synthesis with low micromolar to sub-micromolar activities and these compounds successfully abrogated the proliferation of cancer cells in serine free media. The indole amide series reported here represent an important improvement over previously published PHGDH inhibitors as they are markedly more potent and their mechanism of action is better defined.
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http://dx.doi.org/10.1016/j.bmcl.2019.07.011DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6702104PMC
September 2019

Genetic diversity and haplotype structure of 27 Y-STR loci in a Han population from Jining, Shandong province, eastern China.

Forensic Sci Int Genet 2019 09 14;42:e25-e26. Epub 2019 Jun 14.

Affiliated Hospital of Jining Medical University, Jining, Shandong, PR China.

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http://dx.doi.org/10.1016/j.fsigen.2019.06.008DOI Listing
September 2019

Altered steady state and activity-dependent de novo protein expression in fragile X syndrome.

Nat Commun 2019 04 12;10(1):1710. Epub 2019 Apr 12.

Center for Neural Science, New York University, New York, NY, 10003, USA.

Whether fragile X mental retardation protein (FMRP) target mRNAs and neuronal activity contributing to elevated basal neuronal protein synthesis in fragile X syndrome (FXS) is unclear. Our proteomic experiments reveal that the de novo translational profile in FXS model mice is altered at steady state and in response to metabotropic glutamate receptor (mGluR) stimulation, but the proteins expressed differ under these conditions. Several altered proteins, including Hexokinase 1 and Ras, also are expressed in the blood of FXS model mice and pharmacological treatments previously reported to ameliorate phenotypes modify their abundance in blood. In addition, plasma levels of Hexokinase 1 and Ras differ between FXS patients and healthy volunteers. Our data suggest that brain-based de novo proteomics in FXS model mice can be used to find altered expression of proteins in blood that could serve as disease-state biomarkers in individuals with FXS.
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http://dx.doi.org/10.1038/s41467-019-09553-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6461708PMC
April 2019

Real-Time, in Vivo Correlation of Molecular Structure with Drug Distribution in the Brain Striatum Following Convection Enhanced Delivery.

ACS Chem Neurosci 2019 05 6;10(5):2287-2298. Epub 2019 Mar 6.

Department of Radiology, Molecular Imaging Innovations Institute , Weill Cornell Medicine , New York , New York 10065 , United States.

The blood-brain barrier (BBB) represents a major obstacle in delivering therapeutics to brain lesions. Convection-enhanced delivery (CED), a method that bypasses the BBB through direct, cannula-mediated drug delivery, is one solution to maintaining increased, effective drug concentration at these lesions. CED was recently proven safe in a phase I clinical trial against diffuse intrinsic pontine glioma (DIPG), a childhood cancer. Unfortunately, the exact relationship between drug size, charge, and pharmacokinetic behavior in the brain parenchyma are difficult to observe in vivo. PET imaging of CED-delivered agents allows us to determine these relationships. In this study, we label different modifications of the PDGFRA inhibitor dasatinib with fluorine-18 or via a nanofiber-zirconium-89 system so that the effect of drug structure on post-CED behavior can accurately be tracked in vivo, via PET. Relatively unchanged bioactivity is confirmed in patient- and animal-model-derived cell lines of DIPG. In naïve mice, significant individual variability in CED drug clearance is observed, highlighting a need to accurately understand drug behavior during clinical translation. Generally, the half-life for a drug to clear from a CED site is short for low molecular weight dasatinib analogs that bare different charge; 1-3 (1, 32.2 min (95% CI: 27.7-37.8), 2, 44.8 min (27.3-80.8), and 3, 71.7 min (48.6-127.6) minutes) and is much longer for a dasatinib-nanofiber conjugate, 5, (42.8-57.0 days). Positron emission tomography allows us to accurately measure the effect of drug size and charge in monitoring real-time drug behavior in the brain parenchyma of live specimens.
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http://dx.doi.org/10.1021/acschemneuro.8b00607DOI Listing
May 2019

A carbazole-functionalized metal-organic framework for efficient detection of antibiotics, pesticides and nitroaromatic compounds.

Dalton Trans 2019 Feb;48(8):2683-2691

Key Laboratory of Material Chemistry for Energy Conversion and Storage, Ministry of Education, Hubei Key Laboratory of Materials Chemistry and Service Failure, School of Chemistry and Chemical Engineering, Huazhong University of Science and Technology, Wuhan 430074, P.R. China.

Organic pollutants, such as antibiotics, pesticides, and nitroaromatic compounds (NACs), have posed a great threat to human health and sustainable development. Therefore, the detection of these organic pollutants is of great importance but challenging. In this work, we synthesized a rigid conjugated tricarboxylic acid ligand 4,4'-(9-(4'-carboxy-[1,1'-biphenyl]-4-yl)-9H-carbazole-3,6-diyl)dibenzoic acid (H3CBCD), and employed this ligand to react with Cd(ii) ions to construct a Cd-LMOF, namely [Cd3(CBCD)2(DMA)4(H2O)2]·10DMA (Cd-CBCD). Cd-CBCD features a three-dimensional (3D) supramolecular framework based on two-dimensional (2D) layer structures through ππ stacking interactions. The fluorescence sensing measurements demonstrate that Cd-CBCD can detect nitrofurans (NFs), 4-nitroaniline (4-NA) and 2,6-dichloro-4-nitroaniline (DCN) with high selectivity and sensitivity. This work represents the first carbazole-functionalized metal-organic framework as a fluorescent sensor for the highly efficient detection of antibiotics, pesticides and nitroaromatic compounds.
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http://dx.doi.org/10.1039/c8dt04558eDOI Listing
February 2019

Rac-Mediated Macropinocytosis of Extracellular Protein Promotes Glucose Independence in Non-Small Cell Lung Cancer.

Cancers (Basel) 2019 Jan 2;11(1). Epub 2019 Jan 2.

Meyer Cancer Center, Weill Cornell Medicine-New York Presbyterian Hospital, New York, NY 10021, USA.

Cancer cells can adapt to nutrient poor conditions by rewiring their metabolism and using alternate fuel sources. Identifying these adaptive metabolic pathways may provide novel targets for cancer therapy. Here, we identify a subset of non-small cell lung cancer (NSCLC) cell lines that survive in the absence of glucose by internalizing and metabolizing extracellular protein via macropinocytosis. Macropinocytosis is increased in these glucose independent cells, and is regulated by phosphoinositide 3-kinase (PI3K) activation of Rac-Pak signaling. Furthermore, inhibition of Rac-dependent macropinocytosis blocks glucose-independent proliferation. We find that degradation of internalized protein produces amino acids, including alanine, which generates TCA cycle and glycolytic intermediates in the absence of glucose. In this process, the conversion of alanine to pyruvate by alanine transaminase 2 (ALT2) is critical for survival during glucose starvation. Collectively, Rac driven macropinocytosis of extracellular protein is an adaptive metabolic pathway used by a subset of lung cancers to survive states of glucose deprivation, and may serve as a potential drug target for cancer therapy.
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http://dx.doi.org/10.3390/cancers11010037DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6356657PMC
January 2019

Mitochondrial One-Carbon Pathway Supports Cytosolic Folate Integrity in Cancer Cells.

Cell 2018 11;175(6):1546-1560.e17

Department of Medicine, Meyer Cancer Center, Weill Cornell Medicine, New York, NY 10065, USA. Electronic address:

Mammalian folate metabolism is comprised of cytosolic and mitochondrial pathways with nearly identical core reactions, yet the functional advantages of such an organization are not well understood. Using genome-editing and biochemical approaches, we find that ablating folate metabolism in the mitochondria of mammalian cell lines results in folate degradation in the cytosol. Mechanistically, we show that QDPR, an enzyme in tetrahydrobiopterin metabolism, moonlights to repair oxidative damage to tetrahydrofolate (THF). This repair capacity is overwhelmed when cytosolic THF hyperaccumulates in the absence of mitochondrially produced formate, leading to THF degradation. Unexpectedly, we also find that the classic antifolate methotrexate, by inhibiting its well-known target DHFR, causes even more extensive folate degradation in nearly all tested cancer cell lines. These findings shed light on design features of folate metabolism, provide a biochemical basis for clinically observed folate deficiency in QDPR-deficient patients, and reveal a hitherto unknown and unexplored cellular effect of methotrexate.
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http://dx.doi.org/10.1016/j.cell.2018.09.041DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6568313PMC
November 2018

A Novel Magnesium Metal-Organic Framework as a Multiresponsive Luminescent Sensor for Fe(III) Ions, Pesticides, and Antibiotics with High Selectivity and Sensitivity.

Inorg Chem 2018 Nov 18;57(21):13330-13340. Epub 2018 Oct 18.

Key Laboratory of Material Chemistry for Energy Conversion and Storage, Ministry of Education, Hubei Key Laboratory of Materials Chemistry and Service Failure, School of Chemistry and Chemical Engineering , Huazhong University of Science and Technology , Wuhan 430074 , P. R. China.

Iron(III) ions play a vital role in living biological systems, while organic pollutants including pesticides and antibiotics pose a great threat to the ecological environment. Effective detection for these species is crucial for human health and environmental protection. In this work, we designed and synthesized a new amino-decorated bridging ligand HAPDA and employed it to react with the environmentally friendly Mg(II) ions to construct a novel magnesium luminescent metal-organic framework (Mg-LMOF), namely [Mg(APDA)(HO)]·5DMA·5HO (Mg-APDA). The as-synthesized Mg-LMOF is a three-dimensional framework with one-dimensional hexagonal channels. These microporous channels are decorated with Lewis-base amino sites and uncoordinated O atoms, which facilitate the Mg-APDA to anchor and recognize various analytes. Mg-APDA can be used as a multiresponsive luminescent sensor to detect Fe(III) ions, pesticides, and antibiotics effectively. To the best of our knowledge, this work represents the first amino-decorated Mg-LMOF as an efficient fluorescent sensor for detecting metal ions, pesticides, and antibiotics simultaneously.
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http://dx.doi.org/10.1021/acs.inorgchem.8b01903DOI Listing
November 2018

The vimentin intermediate filament network restrains regulatory T cell suppression of graft-versus-host disease.

J Clin Invest 2018 10 14;128(10):4604-4621. Epub 2018 Aug 14.

Division of Blood and Marrow Transplantation, Department of Pediatrics, Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota, USA.

Regulatory T cells (Tregs) are critical for maintaining immune homeostasis. However, current Treg immunotherapies do not optimally treat inflammatory diseases in patients. Understanding the cellular processes that control Treg function may allow for the augmentation of therapeutic efficacy. In contrast to activated conventional T cells, in which protein kinase C-θ (PKC-θ) localizes to the contact point between T cells and antigen-presenting cells, in human and mouse Tregs, PKC-θ localizes to the opposite end of the cell in the distal pole complex (DPC). Here, using a phosphoproteomic screen, we identified the intermediate filament vimentin as a PKC-θ phospho target and show that vimentin forms a DPC superstructure on which PKC-θ accumulates. Treatment of mouse Tregs with either a clinically relevant PKC-θ inhibitor or vimentin siRNA disrupted vimentin and enhanced Treg metabolic and suppressive activity. Moreover, vimentin-disrupted mouse Tregs were significantly better than controls at suppressing alloreactive T cell priming in graft-versus-host disease (GVHD) and GVHD lethality, using a complete MHC-mismatch mouse model of acute GVHD (C57BL/6 donor into BALB/c host). Interestingly, vimentin disruption augmented the suppressor function of PKC-θ-deficient mouse Tregs. This suggests that enhanced Treg activity after PKC-θ inhibition is secondary to effects on vimentin, not just PKC-θ kinase activity inhibition. Our data demonstrate that vimentin is a key metabolic and functional controller of Treg activity and provide proof of principle that disruption of vimentin is a feasible, translationally relevant method to enhance Treg potency.
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http://dx.doi.org/10.1172/JCI95713DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6159973PMC
October 2018

CRMP4a suppresses cell motility by sequestering RhoA activity in prostate cancer cells.

Cancer Biol Ther 2018 6;19(12):1193-1203. Epub 2018 Aug 6.

b Department of Urology , The University of Kansas Medical Center , Kansas City , KS , USA.

: Distant metastasis is a critical factor for cancer-associated death. Our previous studies identified collapsin response mediator protein 4a (CRMP4a) as a metastasis suppressor in prostate cancers. Enhancing CRMP4 expression by promoter-targeted small activating RNAs reduced cell migration in vitro and abolished distal metastasis in mouse xenograft models. In this study, we investigated the mechanism for CRMP4a-mediated suppression of cell migration. : PC-3 cells were stably infected with lentiviruses expressing CRMP4a cDNA or a shRNA sequence. Cytoskeletal organization was analyzed by measuring cellular focal adhesion area and number, percentage of cell area and lamellipodia numbers after phalloidin staining or anti-vinculin immunocytofluorescent staining. Cell migration was evaluated with Transwell chambers coated with MatriGel. RhoA activation was determined with a Rhotekin RBD agarose bead-based assay kit. Lentiviruses harboring RhoA-Q63L or RhoA-T19N mutant constructs were used to overexpress mutant RhoA proteins. : CRMP4a overexpression largely reduced while CRMP4a knockdown remarkably increased cytoskeletal organization in PC-3 cells. CRMP4a immunoprecipitation pulled down RhoA but not cdc42 or Rac1 proteins. Manipulating CRMP4a expression levels reversely altered active RhoA levels. Overexpression of RhoA active (Q63L) but not inactive (T19N) mutants reversed CRMP4a-mediated reduction of cancer cell migration while RhoA inhibitor Rhosin diminished CRMP4a shRNA-induced increase of cancer cell migration. CRMP4a overexpression also largely reduced cell spreading that was abolished by overexpressing RhoA active mutant. : Our data demonstrated that CRMP4a interacts with RhoA and sequesters its activity, resulting in suppression of cytoskeletal organization, cell migration and spreading.
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http://dx.doi.org/10.1080/15384047.2018.1491507DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6301799PMC
August 2018

CD51 is an independent unfavorable prognostic factor in esophageal squamous cell carcinoma.

Pathol Res Pract 2018 Sep 11;214(9):1352-1355. Epub 2018 Jul 11.

Department of Gastroenterology, Shandong Control Center for Digestive Diseases, Jining, Shandong, 272067, PR China. Electronic address:

Background: Esophageal squamous cell carcinoma (ESCC) is a common cancer in East Asia and some other parts of the world with a dismal prognosis. CD51 (integrin αv),a transmembrane glycoprotein responsible for cell-to-matrix binding has been found to enhance tumor progression. However, its expression and clinicopathological significance in ESCC tumors are not fully understood. The purpose of this study was to investigate the expression level of CD51 and to explore its clinicopathological significance in ESCC.

Methods: The expression of CD51 in 122 ESCC samples was examined by immunohistochemistry and its clinicopathological significance was evaluated.

Results: The expression of CD51 was observed in tumor cell membrane and/or cytoplasm, with a positive rate of 48.36% (59/122). High expression of CD51 was significantly associated with lymph node metastasis (P =  0.031), tumor size (P =  0.028) and invasive depth (P =  0.027). Kaplan-Meier analysis revealed that positive expression of CD51 was correlated with poor overall survival of ESCC patients (P =  0.015). Multivariate analysis suggested that CD51 was an independent prognositic factor for ESCC (hazard ration = 1.604; 95% CI, 1.086-2.368; P =  0.017).

Conclusion: These data suggested CD51 was a predictor for the prognosis of ESCC patients.
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http://dx.doi.org/10.1016/j.prp.2018.07.005DOI Listing
September 2018

Function of Axl receptor tyrosine kinase in non-small cell lung cancer.

Oncol Lett 2018 Mar 27;15(3):2726-2734. Epub 2017 Dec 27.

Cancer Pathology Research Institute, Jining Medical University, Jining, Shandong 272067, P.R. China.

Axl receptor tyrosine kinase (hereafter Axl) is a member of the tyrosine-protein kinase receptor Tyro3, Axl and proto-oncogene tyrosine-protein kinase Mer family of receptor tyrosine kinases, possessing multiple different functions in normal cells. Axl is overexpressed and activated in numerous different human cancer types, triggering several signaling pathways and enhancing tumor progression. The present review assesses previous studies on the function of Axl in non-small cell lung cancer (NSCLC). Axl is overexpressed in the tumor tissues of a number of patients with NSCLC and is associated with poorer clinical outcomes; it promotes NSCLC tumor growth, invasion/metastasis, drug resistance and the epithelial-mesenchymal transition, thus providing a survival advantage to tumor cells. Therefore, Axl may be a promising target in NSCLC treatment.
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http://dx.doi.org/10.3892/ol.2017.7694DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5778882PMC
March 2018

AXL is a marker for epithelial-mesenchymal transition in esophageal squamous cell carcinoma.

Oncol Lett 2018 Feb 20;15(2):1900-1906. Epub 2017 Nov 20.

Cancer Pathology Research Institute, Jining Medical University, Jining, Shandong 272067, P.R. China.

Esophageal squamous cell carcinoma (ESCC) is a common cancer in China and certain other parts of the world with a dismal prognosis for affected patients. AXL is a member of the TYRO3-AXL-MER family of receptor tyrosine kinases, and has been revealed to be an important mediator of epithelial-mesenchymal transition (EMT) in several types of cancer. However, to the best of our knowledge, its function in EMT in ESCC cells has not yet been examined. The present study employed two independent ESCC mRNA profile datasets and revealed that AXL is associated with several EMT markers. Gene Set Enrichment Analysis indicated that EMT occurs more in ESCC with high AXL expression. Analysis on another dataset demonstrated further that increased expression of AXL in ESCC is associated with increased migratory ability. Collectively, the results of the present study provide evidence that AXL is a marker for EMT in ESCC.
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http://dx.doi.org/10.3892/ol.2017.7443DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5774429PMC
February 2018

Metabolomic Analysis of Glioma Cells Using Nanoflow Liquid Chromatography-Tandem Mass Spectrometry.

Methods Mol Biol 2018 ;1741:125-134

Department of Cell Biology, Kimmel Center for Biology and Medicine at the Skirball Institute, New York University School of Medicine, New York, NY, USA.

Mass spectrometry (MS)-based techniques have been finding utility as sensitive, high throughput metabolite analysis tools for complex biological samples. We describe here a nanoflow liquid chromatography-tandem mass spectrometry (nano-LC-MS/MS) system we developed and applied to metabolic profiling of human cells. Metabolites are extracted from cells using methanol, and filtered through a C18 StageTip to remove large particles. Metabolite samples are separated by HPLC at a flow rate of 400-500 nl/min, then analyzed in both positive and negative ion modes in an LTQ-Orbitrap MS. Metabolite identification and differential analysis are performed using commercial or open source software. Protocols outlined in this chapter describe how nano-LC-MS can be applied to investigate metabolic profiling with limited biomass amount.
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http://dx.doi.org/10.1007/978-1-4939-7659-1_10DOI Listing
December 2018

Fenofibrate prevents skeletal muscle loss in mice with lung cancer.

Proc Natl Acad Sci U S A 2018 01 8;115(4):E743-E752. Epub 2018 Jan 8.

Meyer Cancer Center, Weill Cornell Medicine, New York, NY 10021;

The cancer anorexia cachexia syndrome is a systemic metabolic disorder characterized by the catabolism of stored nutrients in skeletal muscle and adipose tissue that is particularly prevalent in nonsmall cell lung cancer (NSCLC). Loss of skeletal muscle results in functional impairments and increased mortality. The aim of the present study was to characterize the changes in systemic metabolism in a genetically engineered mouse model of NSCLC. We show that a portion of these animals develop loss of skeletal muscle, loss of adipose tissue, and increased inflammatory markers mirroring the human cachexia syndrome. Using noncachexic and fasted animals as controls, we report a unique cachexia metabolite phenotype that includes the loss of peroxisome proliferator-activated receptor-α (PPARα) -dependent ketone production by the liver. In this setting, glucocorticoid levels rise and correlate with skeletal muscle degradation and hepatic markers of gluconeogenesis. Restoring ketone production using the PPARα agonist, fenofibrate, prevents the loss of skeletal muscle mass and body weight. These results demonstrate how targeting hepatic metabolism can prevent muscle wasting in lung cancer, and provide evidence for a therapeutic strategy.
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http://dx.doi.org/10.1073/pnas.1714703115DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5789923PMC
January 2018

Low-Grade Astrocytoma Mutations in IDH1, P53, and ATRX Cooperate to Block Differentiation of Human Neural Stem Cells via Repression of SOX2.

Cell Rep 2017 Oct;21(5):1267-1280

Department of Neurosurgery, NYU School of Medicine, New York, NY 10016, USA; Kimmel Center for Stem Cell Biology, NYU School of Medicine, New York, NY 10016, USA; Laura and Isaac Perlmutter Cancer Center, NYU School of Medicine, New York, NY 10016, USA; Brain Tumor Center, NYU School of Medicine, New York, NY 10016, USA; Neuroscience Institute, NYU School of Medicine, New York, NY 10016, USA. Electronic address:

Low-grade astrocytomas (LGAs) carry neomorphic mutations in isocitrate dehydrogenase (IDH) concurrently with P53 and ATRX loss. To model LGA formation, we introduced R132H IDH1, P53 shRNA, and ATRX shRNA into human neural stem cells (NSCs). These oncogenic hits blocked NSC differentiation, increased invasiveness in vivo, and led to a DNA methylation and transcriptional profile resembling IDH1 mutant human LGAs. The differentiation block was caused by transcriptional silencing of the transcription factor SOX2 secondary to disassociation of its promoter from a putative enhancer. This occurred because of reduced binding of the chromatin organizer CTCF to its DNA motifs and disrupted chromatin looping. Our human model of IDH mutant LGA formation implicates impaired NSC differentiation because of repression of SOX2 as an early driver of gliomagenesis.
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http://dx.doi.org/10.1016/j.celrep.2017.10.009DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5687844PMC
October 2017