bluuebunny/crossref_metadata_2025_split huggingface dataset:
New Tricks for an Old Drug: Inhibition of Glyceraldehyde‐3‐Phosphate Dehydrogenase by Dithiolethiones to Suppress Cancer Cell Proliferation
Authors: White Michael R , Miller Thomas W , Switzer Christopher , Chavis John , Xu Pauline , Roberts David , Wink David , Garcin Elsa D 
Published: November 2023

Dithiolethione compounds, such as the well‐characterized chemotherapeutic oltipraz, possess chemopreventive, cytoprotective, anti‐angiogenic, and chemotherapeutic properties. The chemopreventive effects of these compounds are mediated in part by the activation of antioxidant response elements and the increased expression of the tumor repressor PP2a and other cancer‐preventive proteins. Much of the transcriptional control of this response occurs via modification of thiol residues in the Keap1‐Nrf2 transcription factor complex. The 1,2‐dithiole‐3‐thione compound, ACS‐1, has also been shown to inhibit NF‐kB and downstream inflammatory molecules that promote tumor progression. As a result, these compounds can potentially be considered as promising multi‐targeting anticancer drugs.In addition to these established roles for dithiolethione compounds, we show here that ACS‐1 plays a novel role in inhibiting glycolysis, important for survival and proliferation of cancer cells. Here we show that ACS‐1 inhibits glycolysis in human tumor cell lines by targeting the glycolytic enzyme glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH). To understand the mechanism of action of ACS‐1, we have combined cell‐based assays, mutational analysis, kinetic assays, mass spectrometry, and x‐ray crystallography. We propose a mechanism whereby ACS‐1 inhibits glycolysis via covalent modification of GAPDH. Our results suggest that inhibition of GAPDH glycolytic function can be exploited in synergy with other cytotoxic chemotherapies to treat cancer. Cancer cells usually exhibit increased glycolysis (Warburg effect) in order to meet cellular energy requirements. Increasing evidence suggests that inhibition of glycolysis may be an effective strategy in chemoprevention. As such, these studies may provide new avenues for the treatment of cancer with the dithiolethione class of molecules.Support or Funding InformationNational Institutes of Health Grant T32 GM066706 (MRW)
Chemoenzymatic Synthesis of a Novel Sugar Scaffold for Targeting Ligand Development
Authors: Erickson Katelyn M , Hazel Joseph M , Troutman Jerry M 
Published: November 2023

The capsular polysaccharides (CPSs) that encapsulate bacteria aids in overall virulence of the microbe. The bacterial capsules differ across species of bacteria from benign, pathogenic, or symbiotic bacteria. Specific targeting of the surface of the differing organisms would be advantageous in order to select certain types of bacteria over others, for either targeting or detection purposes.In our lab, we have developed a new strategy to specifically target surfaces of certain types of bacteria. The developed targeting strategy is based on the surface coating of Vibrio vulnificus, and specifically the rare N‐acetyl‐quinovosamine (QuiNAc) sugar contained in the sugar polymer, as well as other sugars from the V. vulnificus polysaccharide biosynthesis pathway. The targeting strategy involves naturally occurring enzyme sugar products, as well as an isoprenoid anchor analogue that was developed for use with elucidation of polysaccharide biosynthesis pathways in our lab.Determination of the sugar linkage to the isoprenoid anchor has been analyzed by RP‐HPLC with the use of a modified fluorescent isoprenoid analogue. A further developed analogue has been synthesized in our lab, which contains a different reactive group. This reactive group is used in the development of the targeting ligand for the rare QuiNAc sugar. Following the development of the targeting ligand associated with the rare sugars on specific types of bacteria, the use of the targeting agent is useful in applications of bacterial detection or in antibacterial schemes.Support or Funding InformationNational Institutes of Health AREA Grants R15GM100402 (J.M.T.), R15GM114773 (J.M.T.) and National Science Foundation Instrumentation Grant 1337873 (UNC‐Charlotte Chemistry)
ABO Blood Type Correlates with Survival on Prostate Cancer Vaccine Therapy
Authors: Gildersleeve Jeff 
Published: November 2023

Cancer vaccines and other immunotherapies have significant potential for treating cancer patients, but clinical responses vary considerably from patient to patient. Simple, inexpensive strategies to target treatment to likely responders could substantially improve efficacy while simultaneously reducing health care costs, but identification of reliable biomarkers has proven challenging. Previously, we found that pre‐treatment serum IgM to blood group A (BG‐A) correlated with survival for patients treated with PROSTVAC‐VF, a therapeutic cancer vaccine in phase III clinical trials for the treatment of prostate cancer. These results suggested that ABO blood type might influence efficacy. Unfortunately, blood types were not available in the clinical records for all but 8 patients and insufficient amounts of sera were left for standard blood typing methods. To test the hypothesis, therefore, we developed a new glycan microarray‐based method for determining ABO blood type. The method requires only 4 μL of serum, provides 97% accuracy, and allows simultaneous profiling of many other serum anti‐glycan antibodies. After validation with 220 healthy subjects of known blood type, the method was then applied to 74 PROSTVAC‐VF patients and 37 control patients from a phase II trial. In this retrospective study, we found that type B and O PROSTVAC‐VF patients demonstrated markedly improved clinical outcomes relative to A and AB patients, including longer median survival, longer median survival relative to Halabi predicted survival, and improved overall survival via Kaplan‐Meier survival analysis (p=0.006). Consequently, blood type may provide an inexpensive screen to pre‐select patients likely to benefit from PROSTVAC‐VF therapy.Support or Funding InformationThis research was supported by the intramural research program of the NCI/NIH.
High Affinity Binding of a MUC1 Specific Antibody to a Glycopeptide Mediated by Conformational Selection
Authors: Brooks Cory L , Movahedin Reza , Brooks Teresa M , Supekar Nitin , Boons Geert‐Jan 
Published: November 2023

MUC1 is a transmembrane glycoprotein and is a target for antibody‐mediated immunotherapy. In cancer cells, MUC1 is upregulated, and undergoes abnormal glycosylation resulting in truncated carbohydrate chains being added to the protein. The truncated glycosylation exposes cryptic peptide epitopes that can be recognized by antibodies. Since these immunogenic regions are cancer specific, they represent ideal targets for therapeutic antibodies. The peptide sequence, and the cancer specific truncated glycans are both available for antibody recognition. There is significant interest in determining which is the more suitable target for antibody immunotherapy. We investigated the role of tumor specific glycosylation in antigen recognition by the therapeutic antibody AR20.5. Using microscale thermophoresis we explored the affinity of AR20.5 to cancer specific MUC1 glycopeptide and peptide. The antibody bound to the glycopeptide with an order of magnitude stronger affinity than against the naked peptide. Given these results we postulated that A20.5 must specifically bind the carbohydrate as well as the peptide. Using X‐ray crystallography we examined this hypothesis by determining the structure of AR20.5 in complex with both peptide and glycopeptide. Surprisingly, the structure revealed that the carbohydrate did not form any specific contacts with the antibody. The high affinity of AR20.5 for the glycopeptide and the lack of specific binding contacts supports a hypothesis that glycosylation of MUC1 stabilizes an extended bioactive conformation of the peptide that is recognized by the antibody. High affinity binding of AR20.5 to the MUC1 glycopeptide is not driven by specific antibody‐antigen contacts, but rather glycosylation alters the conformational equilibrium of the antigen which allows the antibody to select the correct coformation. This study demonstrates a novel mechanism of antibody‐antigen interaction and also suggests that glycosylation of MUC1 is important for the generation of high affinity therapeutic antibodies.Support or Funding InformationThis work was supported through start‐up funds from the College of Science and Math at CSUF, and by a research grant from Quest Pharmatech Inc. to CLB. MR was supported by rGrants from ASI at CSUF and FSSRA awards from the College of Science and Math at CSUF.
Longitudinal Monitoring of Nanoparticle Accumulation in PC‐3 Tumors
Authors: Nneji Joseph Chidi , Gharagouzloo Codi Amir 
Published: November 2023

Currently there isn't any definitive and non‐destructive test for predicting the accumulation of nanoparticles within tumors, and longitudinal monitoring of delivery requires a baseline using current methodologies. Diagnostic tools that make quantitative, independent measurements are needed to move product to the market. We utilized a subcutaneous PC‐3 tumor modal in 5 mice to predict and quantify accumulation of ferumoxytol. Diffusion tensor imaging (DTI) was utilized pre‐contrast to map out the diffusion anisotropy on a per‐voxel basis throughout the tumor to generate a predictive map for passive accumulation. QUTE‐CE MRI, a novel imaging modality developed in our lab, along with T1 and T2 measurements was used to follow accumulation.We are developing a methodology for both predicting and unambiguously characterizing the heterogeneous accumulation of nanoparticles within tumors from the enhanced permeability and retention (EPR) effect. This impacts the market of cancer pharmaceuticals, currently limited by its predictive ability of exclusive drug delivery to the tumor.We accurately predicted the heterogeneous accumulation of ferumoxytol with DTI. QUTE‐CE MRI proved to be more efficacious than T1 or T2 measurements, producing a better or comparable CNR, as well as clear, unambiguous definition of uptake without the need for comparison to pre‐contrast images. We have developed a novel strategy for predicting and following the accumulation of nanoparticles within the tumor. Ferumoxytol may be used as surrogate, combined with our methodology, to predict the EPR effect of other nanoparticles for image guided drug delivery.
Claudin‐7 regulates survival and glucose metabolism of human lung cancer cells
Authors: Kim Do Hyung , Jackson Spencer M , Chen Yan‐Hua , Lu Qun 
Published: November 2023

Claudins are a family of tight junction membrane proteins engaged in a variety of human diseases including cancer. It has been reported that lung cancer patients with high claudin‐7 expression showed better survival than those with low claudin‐7 expression. Our study has shown that suppressing claudin‐7 expression (KD) in a human lung adenocarcinoma HCC827 cell line significantly increased cell proliferation in culture and tumor growth in mice. To investigate how tumor microenvironment plays a role in claudin‐7 regulation of cell survival, we treated the control and claudin‐7 KD cells under the hypoxia (1% O2) condition for 3 days. Light microscopic study revealed KD cells with fairly‐normal looking morphology whereas control cells displayed shrunken cell morphology. Immunofluorescence staining confirmed substantially decreased expression of cleaved PARP in KD cells than that of control cells (p < 0.01). To examine how claudin‐7 modulates glucose metabolism for cell survival, the cells were pre‐treated with normal (2g/L) glucose‐containing medium in normal oxygen condition for 2 days, followed by being sub‐cultured with normal or low (0.2g/L) glucose‐containing medium in either hypoxia or normal oxygen condition up to additional 24 hours. Western blot revealed an elevation in phosphorylation (or inhibition) of a negative regulator of glycogen synthesis GSK3β, phospho‐glycogen synthase kinase 3 beta (p‐GSK3β), and a substantial decrease in a cellular energy sensor, phospho‐AMP kinase (p‐AMPK), in KD cells when compared to control cells after the cells were treated in low glucose‐containing media for 24 hours, indicating the enhanced glycogen synthesis in KD cells for cell survival. This observation was not evident when the cells were cultured in the normal oxygen condition. We propose that claudin‐7 participates in regulating glucose metabolism and cell survival in human lung cancer cells through modulating GSK3β and AMPK signaling pathways.Support or Funding InformationThis study was supported by National Institute of Health grants HL085752.
A Novel Role for the Extracellular Matrix in Regulation of Hepcidin in Breast Cancer Spheroids
Authors: Blanchette Nicole , Tesfay Lia , Kita Daniel , Claffey Kevin P , Torti Frank M , Torti Suzy V 
Published: November 2023

Iron is an essential nutrient for breast cancer cell growth and proliferation. Our laboratory has demonstrated that ferroportin (Fpn), an iron efflux pump, plays a critical role in maintenance of intracellular iron levels. Fpn is controlled by hepcidin, a secreted peptide hormone that binds to Fpn and triggers its degradation. We have observed that Fpn expression is down‐regulated and hepcidin expression is up‐regulated in breast cancer cells, promoting increased accumulation of iron in cancer cells to fuel their proliferative demands. Since expression of Fpn/hepcidin predicts outcome in breast cancer patients, we explored pathways that control these proteins. We used a 3D spheroid culture system to model in vivo breast tumor architecture, microenvironment and metabolism. We found that hepcidin expression is significantly up‐regulated from 2D to 3D culture of MCF‐7 and patient‐derived breast cancer cells, suggesting that hepcidin may be regulated by processes that exceed the realm of 2D culture. Culturing MCF‐7 cells as spheroids in the presence of Matrigel repressed the induction of hepcidin, suggesting that exogenous extracellular matrix (ECM) proteins present in Matrigel may negatively regulate hepcidin. Further analysis of the effect of exogenous ECM on hepcidin expression revealed that collagen IV, one of the most abundant basement membrane (BM) proteins in Matrigel, negatively regulates hepcidin expression in MCF‐7 spheroids. On the other hand, endogenous production of ECM components, such as collagen I and collagen II, matrix components frequently up‐regulated in breast cancer, was increased in MCF‐7 spheroids compared to monolayer culture. It is known that there is a modified microenvironement during breast tumorigenesis that includes breakdown of the structural BM proteins and deregulation of matrix components, including those that support tumor progression and promote metastatic spread. Together, our data suggest that components of the ECM play a dynamic role in both positive and negative regulation of hepcidin expression in breast cancer spheroids. This finding suggests that the ECM is an integral component in regulation of hepcidin within the tumor microenvironment. Understanding mechanisms of hepcidin regulation by ECM components will provide a greater understanding of the crosstalk between the ECM and iron metabolism in breast cancer.Support or Funding InformationThis research is supported by NIH R01 CA171101 and NIH R01 CA188025.
Upregulated Expression of GRP78 Supports ER Stress‐induced Apoptosis in Tunicamycin Treated Triple Negative Breast Tumor
Authors: Banerjee Dipak K , Banerjee Aditi , Zhang Zhenbo , Serrano Jesus E , Romero Eva C , Sanchez Neysharie , Baksi Krishna 
Published: November 2023

Breast cancer affects nearly 1.5 million women (5–10%) worldwide per year. Triple negative breast cancer (TNBC) accounts for 15% of all breast cancer with a disproportionate share of mortality. The patients are younger and pre‐menopausal. The cancers are poorly differentiated, and most fall into the basal subgroup of breast cancers. The long‐term survival is extremely low because specific treatment guidelines are lacking. The patients are managed with standard treatment causing a high rate of local and systemic relapse. This make them resistant to existing targeted therapies (endocrine/biologics/adjuvant/neo‐adjuvant). Anthracycline/taxane combination therapy and PARP inhibitor are currently being explored but not known if they would achieve a pathological complete response (pCR). Protein glycosylation has been claimed as a universal feature of helping cancer cells escaping immune surveillance, facilitate tumor invasion, and increase malignancy with increased tumor burden and poor prognosis as well as enhanced angiogenesis. Based on the glycomics profile of human breast cancer cells as well as tumor specimen, we have hypothesized that targeting asparagine‐linked (N‐linked) protein glycosylation would evolve a new generation therapeutic preventing breast tumor progression and eliminating the disease. To test our hypothesis, we have used tunicamycin a natural product that competitively inhibits the N‐acetyl glucosaminyl 1‐phosphate transferase activity in the ER and consequently the N‐linked protein glycosylation. When tested in a triple negative breast tumor in nude mice, the tumor progression is inhibited by ~65% in one week at tunicamycin concentration of 0.25 mg/Kg administered orally twice a week. Human triple negative breast cancer cells (MDA‐MB‐231) are also equally susceptible to tunicamycin action. The cells are arrested in G1 and exhibit ER stress followed by induction of apoptosis mediated by unfolded protein response (upr) signaling. Western blotting and qRT‐PCR support increased expression of the ER stress master regulator GRP78. But, the immunofluorescence microscopy could not detect GRP78 on the surface of control or tunicamycin treated cells. The result is the same in cells cultured in the absence of serum. GRP78 fluorescence however, is detectable in cells after either they are fixed with ice‐cold methanol or permeabilized with digitonin. We, therefore, conclude that GRP78 is not expressed on the outer‐leaflet of the cell surface of the triple negative human breast cancer cells MDA‐MB‐231. But, its intracellular expression is anti‐tumorigenic.Support or Funding InformationSupported in part by NIH U54‐CA096297 and Komen for the Cure BCTR06582 (DKB) and NIH/NIMHD 2G12MD007583 (KB).
Getting the Shaft?: Investigating Midshaft Location in Immature Femora
Authors: Eleazer Courtney D. , Kelso Rebecca Scopa , West Frankie L. , Williams Laura , Stradleigh Ryan , Shaeffer Ari 
Published: November 2023

In skeletal samples, diaphyseal breadths of immature femora have traditionally been measured at 50% of diaphyseal length. However, research on the Denver growth sample suggests that 45.5% of diaphyseal length in younger individuals best represents the most analogous position to the 50% midshaft location in adult femora with fused epiphyses. Currently, direct comparisons have not been made between measures at 45.5% and 50% diaphyseal length. Lack of methodological consistency could affect comparisons across studies of immature skeletal remains.Comparative analysis utilizing modern day radiographs of immature individuals (birth‐12 years) indicated that 50% of diaphyseal length is, on average, located 2.6 mm proximal to true midshaft. This discrepancy is caused by the distal epiphysis contributing more to total bone length than the proximal epiphysis and confirms that mid‐diaphyseal locations approximate, but do not exactly align with, midshaft locations.Additionally, we analyzed immature (birth‐12 years)femora from several archaeological skeletal assemblages (n=170) to determine whether significant differences are present in anterioposterior (AP) and mediolateral (ML) breadths between the two diaphyseal locations. A repeated measures ANOVA indicates significantly wider AP and ML breadths at 45.5% when compared with 50% in all archaeological populations. Additionally, age is positively correlated with increases in AP breadth at 45.5% diaphyseal length. Morphological differences in the two locations support current understanding of shaft development and biomechanical changes with age. This finding, along with results from radiographs, suggests that studies utilizing one location may not be directly comparable to those utilizing the other. Future research will assess the impact of age on midshaft location to determine age‐specific methods for collecting diaphyseal breadths from immature femora.
Mitochondria as signaling organelles in aging
Authors: Titorenko Vladimir , Leonov Anna , Arlia‐Ciommo Anthony , Medkour Younes , Svistkova Veronika 
Published: November 2023

Exogenously added lithocholic bile acid (LCA) is known to extend the chronological lifespan of the yeast Saccharomyces cerevisiae, accumulate in mitochondria and alter mitochondrial membrane lipidome. Using quantitative mass spectrometry, we demonstrated that LCA alters the age‐related dynamics of changes in levels of many mitochondrial proteins, as well as numerous proteins in cellular locations outside of mitochondria. These proteins belong to two regulons, each modulated by a different mitochondrial dysfunction; we call them a partial mitochondrial dysfunction regulon and an oxidative stress regulon. Proteins constituting these regulons (1) can be divided into several “clusters”, each of which denotes a distinct type of partial mitochondrial dysfunction that elicits a different signaling pathway mediated by a discrete set of transcription factors; (2) exhibit three different patterns of the age‐related dynamics of changes in their cellular levels; and (3) are encoded by genes whose expression is regulated by the transcription factors Rtg1p/Rtg2p/Rtg3p, Sfp1p, Aft1p, Yap1p, Msn2p/Msn4p, Skn7p and Hog1p, each of which is essential for longevity extension by LCA. Our findings suggest that LCA‐driven changes in mitochondrial lipidome alter mitochondrial proteome and functionality, thereby enabling mitochondria to operate as signaling organelles that orchestrate an establishment of an anti‐aging transcriptional program for many longevity‐defining nuclear genes. We propose a model for how such LCA‐driven changes early and late in life of chronologically aging yeast cause a stepwise development of an anti‐aging cellular pattern and its maintenance throughout lifespan.Support or Funding InformationSupported by NSERC of Canada.
Methylene Blue Alleviates Nuclear and Mitochondrial Abnormalities in Hutchinson‐Gilford Progeria Syndrome
Authors: Xiong Zhengmei , Choi Ji Young , Wang Kun , Zhang Haoyue , Tariq Zeshan , Wu Di , Cao Kan 
Published: November 2023

Hutchinson‐Gilford progeria syndrome (HGPS), a fatal premature aging disease, is caused by a single nucleotide mutation in the lamin A gene. Previous reports have focused on nuclear phenotypes in HGPS cells, and the potential contribution of the mitochondria, a key player in normal aging, remains unclear. Using high‐resolution microscopy analysis, we demonstrated a significantly increased fraction of swollen and fragmented mitochondria and a marked reduction in mitochondrial mobility in HGPS fibroblast cells. Notably, the expression of PGC‐1α, a central regulator of mitochondrial biogenesis, was inhibited by progerin. To rescue mitochondrial defects, we treated HGPS cells with a mitochondrial‐targeting antioxidant methylene blue (MB). Our analysis indicated that MB treatment not only alleviated the mitochondrial defects but also specifically rescued the hallmark nuclear abnormalities in HGPS cells. Additional analysis suggested that MB treatment released progerin from the nuclear membrane, rescued perinuclear heterochromatin loss and corrected misregulated gene expression in HGPS cells. Together, these results demonstrate a role of mitochondrial dysfunction in developing the premature aging phenotypes in HGPS cells and suggest MB as a promising therapeutic for HGPS.Support or Funding InformationNIH/NHGRI Grants R01HG007104 (KC) and R21AG043801 (KC)