bluuebunny/crossref_metadata_2025_split huggingface dataset:
Drugging the undruggable steroid receptor coactivators
Authors: Wang Jin 
Published: November 2023

Protein–protein interactions (PPIs) play a central role in most biological processes, and therefore represent an important class of targets for therapeutic development. However, disrupting PPIs using small‐molecule inhibitors (SMIs) is challenging and often deemed as “undruggable.” We developed a cell‐based functional assay for high throughput screening to identify SMIs for steroid receptor coactivator‐3 (SRC‐3 or AIB1), a large and mostly unstructured nuclear protein. Without any SRC‐3 structural information, we identified SI‐2 as a highly promising SMI for SRC‐3. SI‐2 meets all of the criteria of Lipinski's rule for a drug‐like molecule and has a half‐life of 1 h in a pharmacokinetics study and a reasonable oral availability in mice. As a SRC‐3 SMI, SI‐2 can selectively reduce the transcriptional activities and the protein concentrations of SRC‐3 in cells through direct physical interactions with SRC‐3, and selectively induce breast cancer cell death with IC50 values in the low nanomolar range (3–20 nM), but not affect normal cell viability. Furthermore, SI‐2 can significantly inhibit primary tumor growth and reduce SRC‐3 protein levels in a breast cancer mouse model. In a toxicology study, SI‐2 caused minimal acute cardiotoxicity based on a hERG channel blocking assay and an unappreciable chronic toxicity to major organs based on histological analyses. We believe that this work could significantly improve breast cancer treatment through the development of “first‐in‐class” drugs that target oncogenic coactivators.Support or Funding InformationThis work is supported in part by National Institutes of Health Grants R01GM115622, Cancer Prevention and Research Institute of Texas Grants R1104, Welch Foundation Grant Q‐1798; the Center for Comparative Medicine, the Cytometry and Cell Sorting Core, the Center for Drug Discovery, and the Dan L. Duncan Cancer Center at Baylor College of Medicine; and the Texas Medical Center Digestive Diseases Center.
Total Synthesis, Antileishmanial and Anticancer Activity of the Acetylenic Fatty acids 6‐Hexadecynoic, 10‐Phenyl‐6‐decynoic, and 10‐Cyclohexyl‐6‐decynoic Acids
Authors: Álvarez Ester , Carballeira Néstor , Morales Christian , Delgado Yamixa , Tinoco Arthur , Reguera Rosa , Álvarez Raquel , Balaña Rafael 
Published: November 2023

Naturally occurring acetylenic fatty acids have been recognized for their biological activity towards cancer and protozoal parasites. Lung cancer is one of the deadliest types of cancer and the second most incidental cancer in both men and women. Previous research has shown that these acids display cytotoxicity against the human lung carcinoma cell line A549. On the other hand, leishmaniasis is a neglected disease caused by the protozoan parasite Leishmania donovani(infantum) that affects mostly underdeveloped countries and can be fatal in the most severe cases (visceral leishmaniasis). Our laboratory has shown that the Δ6 alkynoic fatty acids display antiprotozoal activity and inhibit the Leishmania donovani topoisomerase IB enzyme (LdTopIB). In this work, the total syntheses of the naturally occurring 6‐hexadecynoic acid and the synthetic 10‐phenyl‐6‐decynoic and 10‐cyclohexyl‐6‐decynoic acids were completed in 5 steps (67–77% overall yields). The 6‐hexadecynoic acid displayed good inhibition of LdTopIB (EC‐‐50=61 μM) and cytotoxicity towards L. infantum iRFP70 promastigotes (IC50=100 μM). In addition, the 6‐hexadecynoic acid also displayed cytotoxicity towards the human lung A549 cell line (EC‐50= 65 μM). On the other hand, the 10‐phenyl‐6‐decynoic acid did not inhibit LdTopIB (EC‐‐50>200 μM) and displayed no cytotoxicity towards L. infantum iRFP70 promastigotes (IC50>100 μM) as well as no cytotoxicity towards A549 (EC50>100 μM). The 10‐cyclohexyl‐6‐decynoic acid also showed no inhibition of LdTopIB, no cytotoxicity towards L. infantum promastigotes, but displayed cytotoxicity against the A549 cell line (EC‐50= 87 μM). Our results show that the 6‐hexadecynoic acid was the best of the series due to its inhibitory activity towards LdTopIB and toxicity towards L. infantum promastigotes and the A549 cell line.Support or Funding InformationNIH‐MARC Program undergraduate fellowship (5T34GM00781‐37) and the NIH‐RISE Program graduate fellowship (5R25GM061151‐15)
In vivo drug discovery for progressive supranuclear palsy using a novel zebrafish model
Authors: Burton Edward A , Bai Qing 
Published: November 2023

Progressive supranuclear palsy (PSP) is a common neurodegenerative disorder causing progressive motor, oculomotor, bulbar, and cognitive deficits. Current treatments do not arrest disease progression and the prognosis for functional independence and survival is correspondingly poor. Pathologically, PSP is characterized by accumulation of insoluble hyperphosphorylated forms of the 4‐repeat isoform of the microtubule associated protein tau (4R‐Tau) and subsequent loss of CNS neurons. Convergent lines of evidence suggest that 4R‐Tau is mechanistically involved in the neurodegeneration of PSP. Clinical and pathological PSP phenocopies can result from splice site mutations in the MAPT gene causing over‐expression of 4R‐Tau. Furthermore, PSP is strongly associated with the H1 MAPT haplotype that gives rise to relative over‐production of 4R‐Tau. Consequently, compounds that prevent neurodegeneration caused by 4R‐Tau in vivo may be effective therapeutic agents in PSP.The long‐term goal of our work is to develop effective treatments for progressive supranuclear palsy (PSP). Development of therapies for PSP and other tauopathies has been limited by lack of animal models that allow discovery‐based approaches, for example screening candidate chemical modifiers against disease‐relevant phenotypes in vivo. The zebrafish is a genetically and chemically tractable vertebrate model that is particularly suited to drug discovery and comparative efficacy testing and is being used increasingly for studying human neurological diseases. The objective of this study was to develop a zebrafish model of PSP and to carry out a phenotype based chemical modifier screen in vivo.We generated transgenic zebrafish that express human 4R‐Tau in CNS neurons. Tau transgenic zebrafish replicated many clinic‐pathological features of PSP, including: hypokinesia; oculomotor deficits; somato‐dendritic accumulation of hyperphosphorylated tau; neuronal apoptosis; and impaired survival. Importantly, these phenotypes were quantitative, replicable and occured rapidly allowing their use as endpoints in a phenotype‐driven screen. We tested over 200 bioactive compounds to identify chemicals that rescued motor function in the 4R‐Tau zebrafish, using a custom automated video tracking and motion analysis method. We identified several ‘hits’ that rescued motor function and showed activity in subsequent verification assays.Our completion of the first unbiased phenotype‐based chemical modifier screen in a vertebrate tauopathy model provides proof‐of‐concept that drug discovery can be carried out in vivo by assaying disease‐relevant neurological deficits. Importantly our screen has so far identified several candidates for further development as possible tau therapeutics.Support or Funding InformationUS Department of Veterans' Affairs (BX003168‐01)NINDS (NS080881)CurePSP (441‐05, 468‐08)Pittsburgh Foundation (AD201577642)
The Antimicrobial Property of the Acetone Extract of Cola Acuminate
Authors: Thomas Brianna J , Telles Caroline M 
Published: November 2023

The increase in drug‐resistant bacteria during the recent years has sparked an interest in antimicrobial properties of medicinal plants. Thus, the mechanism by which medicinal plants kill bacteria is certainly an important topic that merits further research. Cola acuminate, also known as Obi or Bizzy nut to the Ettu people of Jamaica, is a “cure‐all” herbal medicine. Cola acuminate (Bizzy Nut), possesses a variety of bioactive compounds which exhibit important inhibitory activities against the growth of certain mammalian cell lines, bacteria and fungi. However, the specific chemicals responsible for this bioactivity have not been identified. Thus, we hypothesized that the solid‐liquid extraction of Bizzy Nut, followed by antimicrobial screening would lead to the identification of active antimicrobial agents present in Bizzy Nut.As an initial step in testing our hypothesis, finely ground samples of Bizzy nut were sequentially extracted in a Soxhlet apparatus using 100% hexane, ether, acetone, methane, or water to produce five independent extracts of unique polarity. The antimicrobial activity of the acetone extract was carried out using Staphylococcus aureus and Pseudomonas aeruginosa as test organisms. Our results show that the acetone extract (Biz‐3) possesses antimicrobial activity in Staphylococcus aureus at concentrations as low as 40ppm. There was a dose‐dependent increase in the zone of inhibition of Biz‐3 from 8.5±0.30mm at 40ppm to 11.7±0.1 mm at 120ppm. To purify this antimicrobial activity, we have used SPE to generate three unique fractions of the acetone extract. The antimicrobial screening (Minimum Inhibitory Concentration, MIC) of each of the SPE fraction will be discussed. Our conclusion from this work is that the acetone extract of Cola acuminate (Bizzy Nut), contains putative antimicrobial bioactivity compounds that are effective again gram positive bacteria.
An investigation of the oxidatively damaged transcriptome in the human neuronal cells
Authors: Kharel Prakash , Gadepalli Venkat , Chattopadhyay Anshuman , McDonough Jennifer , Basu Soumitra 
Published: November 2023

Reactive oxygen species (ROS) are produced intracellularly and can cause oxidative damage to the biomolecules. Under normal physiological conditions ROS are used in cellular homeostasis and cell signaling but during stress condition they are often overproduced. The overproduction of the ROS is associated with age and neurological disorders, such as, Alzheimer's disease, Parkinson's disease and multiple sclerosis. The weakened antioxidant defense mechanism in the brain of the patients with the neurological disorders makes the neurons susceptible to oxidative damage. The ROS can damage every major class of biomolecules including DNA, RNA and proteins. In the past the focus was mainly on studies of DNA and protein damages, while the detrimental effects of RNA damages only began to be highlighted in recent years. 8‐Hydroxyguanosine (8‐OHG) is the most common oxidative modification found in the RNA. By identifying the modified mRNA molecules during oxidative stress in the neuronal cells (SH‐SY5Y), we aim to delineate the relationships between specific mRNA oxidation and neurodegeneration. We have isolated the modified mRNA from the stressed SH‐SY5Y cells by immunoprecipitation with anti 8‐OHG antibody. Using RT‐qPCR assay we have shown the differential mRNA expression during the oxidative stress, for example, high expression of superoxide dismutase 1 and carbonyl reductase mRNAs. Additionally, we have identified the selectively modified mRNA molecules by using iLumina® RNA sequencing platform. This study will shed light on the yet to be understood link between mRNA oxidation and neurodegeneration.Support or Funding InformationNo travel support yet.
Deciphering the Gene Expression Pattern from Human Blood Samples Collected in Multiple Collection Tubes
Authors: Hoke Allison , Gautam Aarti , Donahue Duncan , Miller Stacy Ann , Srinivasan Seshamalini , Detwiler Leanne , Lynch Jesse , Levangie Michael , Sowe Bintu , Hammamieh Rasha , Jett Marti 
Published: November 2023

Theobjective of this study was to investigate how blood storage, extraction methodologies, and the actual blood component itself may influence what genes are expressed in downstream applications. To discover potential biases, blood was collected in triplicate from five donors into RNAgard Blood Tubes, PAXgene Blood RNA Tubes, EDTA Tubes, Sodium Citrate Cell Preparation Tubes (CPT), and Blood Collection Tubes with Acid Citrate Dextrose Solution A (ACD‐A). The RNA was isolated from whole blood and peripheral blood mononuclear cells (PBMCs) using ten different combinations of experimental conditions combined with several widelyutilized RNA isolation methods. After extraction, the RNA was quantified using a spectrophotometer and RNA quality was assessed by the RNA Integrity Number (RIN) value. The RNA was then profiled using Agilent Human Gene Expression micro arrays to observe the expression levels for the different experimental conditions. When the RNA quality was assessed by the RIN value, we found that all PBMC procedures had the highest RIN values as they were processed and stabilized in TRIzol Reagent prior to RNA extraction. Initial data analysis showed that human blood stored at 4°C overnight performed equally well when compared to frozen stabilized blood. We compared within and across donor and method replicates, which indicated signal to noise patterns that were not captured by RIN value alone. Our results provide some new insights into RNA isolation from blood samples and how it is influenced by the choice of storage, handling, and methodologies, and that careful consideration is necessary to avoid bias in downstream processing. Better characterization of collection method idiosyncrasies will facilitate further research in to understanding the effect of gene expression on variability in human sample types.DISCLAIMERS: Research was conducted in compliance with all Federal Requirements. The views expressed are those of the authors and do not constitute endorsement by the U.S. Army.
Dissecting JADE1 pathway in the Regulation of Cytokinesis
Authors: Panchenko Maria V , Shao Bo 
Published: November 2023

Cell cycle regulation is important for normal proliferation of epithelial cells and epithelial tissue homeostasis. The dis‐regulation of the cell cycle progression is linked to genetic instability, aneuploidy, and apoptosis. Cytokinesis, the final stage of cell division, is responsible for the proper partitioning of duplicated chromatin and other cell content between two distinct daughter cells. We previously identified that JADE1 protein associates with chromatin and promotes acetylation of bulk histone H4 by functioning as an adapter and scaffold in the HBO1 complex, which is histone acetyl transferase (HAT) responsible for bulk histone H4 acetylation in vitro and in cultured cells. Our finding of cell cycle mediated JADE1S chromatin shuttling and phosphorylation suggested JADE1 function in cell cycle progression, specifically during G2/M to G1 progression.Objectiveto examine JADE1 role in the regulation of the cell cycle.Materials and methodscultured cell lines, cDNA transfection, transduction, RNA interference, FACS, western blot, fluorescent confocal microscopy.ResultsThe depletion of JADE1 in synchronized dividing cells increased rates of G2/M to G1 progression as evident from kinetics of G1 cell accumulation. Importantly, JADE1S depletion decreased proportion of cytokinetic cells and increased multinucleates, while JADE1 over‐expression increased cytokinetic proportion. Interestingly, endogenous JADE1 in late telophase was associated with midzone and in cytokinesis with midbody. Inhibition of key mitotic regulator Aurora B kinase with a drug released JADE1S' effects on cytokinesis. These data demonstrate role in cytokinesis and suggest role in abscission delay. Further, we investigated whether HAT HBO1 and unresolved chromatin are involved in JADE1 cytokinesis pathway. According to our results, HBO1 exerts antagonistic effects on JADE1 function by interfering with JADE1‐mediated cytokinesis arrest. Moreover, HBO1 over‐expression compromised midbody associations of JADE1 in early cytokinesis.ConclusionsJADE1 represents novel cytokinesis factor; HAT HBO1 interferes with JADE1 function in cytokinesis.Support or Funding InformationNIH RO1 DK087910, Mallory Fund (Boston Medical Center, Pathology)
DETERMINING THE LINK BETWEEN MAMMALIAN p38 STRESS KINASE PATHWAY AND PEROXISOMES
Authors: Singh Rajnish , Chrestensen Carol 
Published: November 2023

The inability of the body to fight oxidative stress is the cornerstone of aging and diseases such as inflammation, cancer, diabetes and Alzheimer's. The response of the cell to oxidative stress involves two key players, the p38 stress kinase pathway and the peroxisome, and the interaction between this pathway and peroxisome is relatively unknown. Several pieces of evidence point to a role for protein kinases in phosphorylating peroxisome proteins and regulating important peroxisome functions during stress. Plant kinases and phosphatases have been shown to localize to the peroxisome under stress conditions. Mammalian ATM kinase, a kinase activated by DNA damage, has also been shown to translocate to peroxisomes, during oxidative stress, phosphorylating and activating proteins that regulate peroxisome turnover. Some peroxisome biogenesis factors in yeast require phosphorylation to function.. This project seeks to determine if the p38 activated Mapkap kinase 2 isoform 2 (MK2) translocates to the peroxisome and serves as the link between the p38 stress kinase pathway and the stress alleviating organelle, the peroxisome. Sequence analysis of human MK2 shows the presence of ‐TRL, a putative peroxisome targeting sequence (PTS1) at its C‐terminus. The consensus peroxisome targeting sequence ‐SKL found on many peroxisome proteins, binds to a cytosolic receptor called pex5, which then targets the protein to the peroxisome. In this study, crude peroxisome fractions were isolated from fresh chicken livers using differential centrifugation and probed with isoform specific MK2 antibodies. Preliminary western blotting experiments indicate the presence of both MK2 isoforms in the crude peroxisome fractions. Additionally, experiments were done to determine the physical interaction between the putative peroxisome targeting sequence in mapkap kinase 2 (‐TRL) and pex5, using the yeast two‐hybrid assay. Initial results indicate that MK2, isoform 2, shows a weak interaction with Pex5 suggesting that MK2 isoform 2 maybe localizing to the peroxisome through its putative peroxisome targeting sequence. The link between mapkap kinase 2, a stress activated protein kinase and a cellular organelle that is important in combating cellular stress is novel and has the potential to lead to exciting new mechanisms in stress signaling.Support or Funding InformationIncentive Funding for Research & Creative Activity, Center for Excellence in Teaching and Learning, Kennesaw State University
Exosomes‐mediated mTOR activation is involved in ovarian cancer cell aggression and migration
Authors: Madigan Audrey Pamela , Gallant Benjamin , Clark Matthew P , Wan Yinsheng , Zhang Wei , Lu Hongguang 
Published: November 2023

Ovarian cancer remains difficult to treat, with the remission rate still high and its drug resistance obvious. Our previous studies have demonstrated that ovarian cancer cells resist chemotherapeutic drugs via activation of EGFR‐PI3K‐AKT mTOR cell survival signaling pathway. Recent studies have suggested that exosome biogenesis and release is associated with cancer cell aggression, migration and cell‐cell communication. We hypothesize that exosomes‐mediated mTOR activity is involved in ovarian cancer cell metastatic potential and migration. Using phagokinetic motility assay, we observed that inhibitors of PI3K/AKT, MEK/ERK, and mTOR reduced cell migration in ovarian cancer cells (CaOV3 cells). Through confocal microscopy and Western Blot we found that when CaOV3 cells are treated with FBS and/or EGF, higher amounts of CD‐63 and p‐S6 are found. We also found that inhibitors of exosome biogenesis and release, GW4869 and Brefeldin A, attenuated EGF‐ and serum‐induced cell proliferation and migration. Western blot analysis showed that EGF induced expression of CD63, one of the markers of exosomes, in a dose dependent manner. It was also seen that Brefeldin A and Monensin reduce the presence of CD‐63 and p‐S6. Interestingly, while Brefeldin A blocked serum‐induced mTOR activation, GW4869 appeared to enhance mTOR activity in CaOV3 cells. Collectively, our data suggests that exosome biogenesis and release feedback regulates mTOR activity in ovarian cancer cells, with a result of aggression, migration, and drug resistance. Modulation of exosome activities may provide clinical therapeutic options for ovarian cancer.Support or Funding InformationRI‐IBRE
Signaling and secretion of chemotactic factors regulated by calcium‐sensing‐receptor mutants found in breast cancer patients
Authors: Orduña Lennis Beatriz , Hidalgo‐Moyle Joseline Janai , Vázquez‐Prado José , Reyes‐Cruz Guadalupe 
Published: November 2023

The calcium‐sensing‐receptor (CaSR) belonging to the class C of the GPCR family regulates cell proliferation, calcium homeostasis and secretion, among other processes, depending on the tissue in which this receptor is expressed. Its physiological importance is also evidenced by several disorders in the calcium homeostasis associated with a number of naturally mutations. The bioinformatic study of CaSR in the catalogue of somatic mutations in cancer (COSMIC) has documented 419 mutations of this receptor in patients with cancer. Sixteen of which are specifically found in breast cancer, four of them (CaSRN639K, CaSRT732A, CaSRR886Q and CaSRV894I) were generated and study in our laboratory to analyze their role in the secretion of chemotactic factors and the signaling events. Considering the potential coupling of CaSR to multiple heterotrimeric G proteins, we examined the effect of PTX (pertussis toxin), an inhibitor of Gi protein; H89, a PKA inhibitor; and U73122, a phospholipase C inhibitor on ERK activation. Our results demonstrated that CaSRWt signals through Gi and Gq proteins, however the mutants receptors were potentially linked to Gs as well as Gq, and the CaSRN639K could also activates Gi. Importantly, our results indicate that the mutant receptors promoted different patterns in the chemotactic factors secretion analyzed by the potential effect of conditioned media obtained from each mutant receptors to promote migration of porcine aortic endothelial cells (PAE) in Boyden chamber assays. The mutants CaSRT732A and CaSRV894I promoted the secretion of chemotactic factors in a calcium‐dependent manner comparable to the CaSRWt although these secretion is minor with respect to the wild type receptor. The mutant CaSRN639K increased the secretion in a calcium‐independent manner, and its stimulation does not have an effect on secretion. Finally, the CaSRR886Q mutant receptor increase the secretion of chemotactic factors independently of stimulus, and surprisingly receptor stimulation inhibited the secretion of chemotactic factors.CONCLUSIONThe calcium sensing receptor mutants found in breast cancer patients potentially change the G‐protein coupling and elicited different secretory patterns, probably due to change in the G‐protein coupling.Support or Funding InformationThis work was supported by Consejo Nacional de Ciencia y Tecnologia (CONACyT) grants 240119 (to G.R.C.) and 152434 (to J.V.P.) and Fundación Miguel Aleman, A. C. (to G.R.C.). Lennis Beatriz Orduña Castillo is graduate student supported by a CONACyT fellowship (scholarship: 455391)