bluuebunny/crossref_metadata_2025_split huggingface dataset:
Novel Rac Inhibitors as Targeted Therapeutics for Metastatic Breast Cancer
Authors: Asencio‐Torres Gabriela , Hernández Eliud , Vlaar Cornelis , Dharmawardhane Suranganie , Castillo‐Pichardo Linette 
Published: June 2021

Metastatic disease is the primary cause of breast cancer mortality, due to the lack of effective therapy. The Rho GTPase Rac is integral for the promotion of cancer cell migration/invasion, proliferation, and survival. Since metastatic breast cancers often overexpress or exhibit high Rac activity, inhibition of Rac is a viable strategy against metastatic cancer. Recently, we characterized EHop‐016, a small molecule that inhibits Rac activity of metastatic breast cancer cells with an IC50 of 1 μM. EHop‐016, which is 10–100 times more active than previously available Rac inhibitors, inhibits the activity of the Rac downstream effector p21 activated kinase (PAK), lamellipodia extension, and cell migration of metastatic breast cancer cells. Moreover, EHop‐016 at ≥ 25 mg/kg Body Weight (BW) significantly reduced tumor growth, metastasis, and angiogenesis in a mouse model. However, our recent pharmacokinetic study of EHop‐016 in a mouse model demonstrated that its bioavailability should to be improved for further pharmacological development. Our hypothesis is that improvement of the EHop‐016 structure will provide probes with increased potency against Rac and improved bioavailability. Herein we have tested several Ehop‐016 derivatives for their effects on breast cancer cell viability, cell migration, and Rac activation. Using MTT assays we found that the Ehop‐016 derivatives, HV‐107 and HV‐118, significantly inhibit the viability of metastatic breast cancer cell lines MDAMB‐231 and MDA‐MB‐435; while presenting minimal toxicity towards non‐cancerous mammary epithelial cells. The effects of HV‐107 and HV‐118 on the inhibition of Rac activation were tested by ELISA‐based Rac activity assays and pulldown assays. Results show that at 250 nM, HV‐107 inhibits Rac activation by 55%, whereas HV‐118 has a similar effect at 100 nM in MDA‐MB‐231 and MDA‐MB‐435 cells. Moreover, using transwell assays we studied the effects of both Rac inhibitors in metastatic breast cancer cell migration and observed a significant reduction. Finally, we tested HV‐107 in a mouse model of metastatic breast cancer and found a 25% reduction in tumor growth in mice treated with 5 mg/kg BW HV‐107. Taken together, our findings suggest HV‐107 and HV‐118 as promising Ehop‐016 derivatives with potential as anti‐metastatic agents, that should be further characterized.Support or Funding InformationThis study was supported by awards from the Susan Komen for the Cure, NIH/NIMHHD U54MD008149, and the Puerto Rico Science and Technology Trust to SD; NIH/NCRR R25GM061838 to UPR MSC; NIH/NIMHHD RCMI 8G12MD007583RCMI, Title V PPOHA 031M10505 and Title V Cooperative P031S130068 from U.S. Department of Education to UCC; and PRINBRE (NIH/NIGMS P20GM103475‐13) Sub‐Award to LCP.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Regulation of Extracellular IGFBP‐3 by Humanin in A549 Cells
Authors: Muterspaugh Robert Dean , Price Deanna , Guthrie Jeffrey , Heyl Deborah , Evans Hedeel 
Published: June 2021

The insulin‐like growth factor binding protein 3 (IGFBP3) is a multifunctional protein that belongs to a family of IGF binding proteins with highly conserved structures. Previously, an inverse relationship was demonstrated between serum or plasma levels of the protein and lung cancer risk. The protein was found to inhibit the growth and survival of non‐small cell lung cancer (NSCLC) A549 cells and its overexpression potently induced apoptosis in these cells. Humanin is a small mitochondrial‐derived peptide known to bind with high affinity to residues 242–259 of IGFBP‐3. Growing evidence suggests that, when secreted, humanin is a potent cytoprotective peptide against a range of disease models and cellular stress. The role that humanin plays in tumorigenesis and metastasis is not clear in A549 cells.While both IGFBP‐3 and humanin are known to exert their function both intracellularly and extracellularly, little is known about their extracellular interaction and regulation of A549 cell survival.Our hypothesis is that humanin acts as an IGFBP‐3 sponge in the extracellular compartment and thereby regulates cell growth and survival in a time and concentration dependent manner. Using peptide synthesis, ELISA, immunoprecipitation, Western blotting, and tissue culture, we examine the physical and functional interaction of extracellular IGFBP3 and humanin both in vitro and in vivo in A549 cells.Support or Funding InformationThis work was supported by an Eastern Michigan University Provost Research Support Award/Chemistry Seller's FundThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Expression and Regulation of CHP and NHE in Cancer
Authors: Bakker Clare , Wallert Mark , Provost Joseph 
Published: June 2021

The hypoxic and serum starved environment of metastasizing cancer cells requires a mechanism to overcome the low intracellular pH induced by the Warburg effect. The sodium hydrogen exchanger isoform 1 (NHE1) is a highly regulated, transmembrane protein that modulates pH of extra and intra cellular environment. One set of proteins which bind and regulate NHE1 include calcineurin homologous proteins 1 and 2 (CHP1&2). CHP1 is necessary for basal NHE1 transport while CHP2, increases the exchange activity and Vmax of NHE1. Both proteins bind to nearly the same NHE1 residues however these interactions are not clearly known. To understand the role that all three proteins may play in regulating cellular function, tissue expressional distribution was conducted on tumor and non‐diseased human tissues. Analysis of RNA sequence from Expression Atlas database revealed that different from published record, CHP2 is expressed within a number non‐diseased tissue including but not limited to: spinal cord, urinary bladder, vagina, tibial nerve, lower leg skin. Analysis of RNA sequence databases of cancerous tissue showed significantly higher expressed CHP2 within a number of, but not all cancer tissues including lung, cervical, and esophogeal cancers. Surprisingly prostate and colorectal adenocarcinoma express CHP2 with a 3–10 fold decrease in expression compared to non‐diseased tissue. To further analyse the expression of CHP2 and NHE1 in lung cancer cells, we performed qPCR on human tissue samples. Cq values displayed a significantly higher expression of CHP2 within squamous cell carcinoma, but not adenocarcinoma, in comparison to adjacent healthy tissue. RNAseq, Expression Atlas, analysis of CHP2 within non‐small cell lung cancer also demonstrates this 15‐fold increased expression for squamous cell carcinoma. This signals an integral role of CHP2 for the survival of cancer cells, and hints at the subsequent impact upon the regulation of NHE1 to allow cells to survive nascent tumour acidification. To examine this possibility, we measured the impact of pHi in several cancer cell lines treated with an NHE1 specific inhibitor or after NHE1 or CHP2 knockdown. This work shows that CHP2 expression is required for cells to maintain sustainable pHi for survival. The impact of NHE1 in hypoxic conditions several tumor cell lines. Thus, the increase in expression and change in transport kinetics demonstrates the effect of CHP2 upon proton transport for NHE1, recovering pHi and increasing cell survival.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Role of mTORC2 and Nitric oxide in Bladder Cancer Invasion
Authors: Sahu Divya , Klemke Richard , Boss Gerry R. , Hansel Donna E. 
Published: June 2021

BackgroundDespite accounting for around 17,000 deaths and 79,000 new diagnoses annually in the United States, treatment options for bladder cancer remain limited. The key driver of bladder cancer stage progression is increasing invasion into the bladder wall, which has limited response to current chemotherapy options and hence newer therapies directed against invasion are key to improving the treatment of bladder cancer. We have recently shown that arginine, the nitric oxide (NO) precursor, is implicated in bladder cancer progression. Our laboratory has also identified mammalian target of rapamycin complex 2 (mTORC2) as a driver of bladder cancer cell invasion. We evaluated the interaction between these two pathways to promote invasion.MethodsExpression of inducible and endothelial nitric oxide synthases (iNOS and eNOS) in FFPE tissue sections of progressive disease was assessed by IHC and correlated with histopathology, progression and stage based on moderate to strong expression (2+, 3+) compared to absent to weak expression (0, 1+). We utilized gene silencing methods and NOS inhibitors and NO scavenger for effects on bladder cancer invasion and migration. We assessed mTORC2 pathway activity and NOS levels in invasive cell tip protrusions called “invadopodia” and evaluated if mTORC2 regulates NOS localization. We used a novel zebrafish model to characterize the effects of mTORC2 and NO on bladder cancer metastases.ResultWe found that eNOS and iNOS are elevated in invasive human bladder tumors and cell lines and their ablation reduces bladder cancer cell migration and invasion by reorganizing the actin cytoskeleton and reducing invadopodia. mTORC2 silencing can affect levels of iNOS and eNOS in bladder cancer cell lines. Silencing of mTORC2, eNOS or iNOS reduced metastases of bladder cancer cells within zebrafish.ConclusionmTORC2 pathway is a key driver of bladder cancer invasion and metastases by regulating the NO pathway and both mTORC2 and NO can be targets for bladder cancer therapy, which would benefit patient outcomes.Support or Funding InformationPrevent Cancer Foundation Fellowship to DSCase Western Reserve University/Cleveland Clinic CTSA Grant Number UL1 RR024989 from the National Center for Research Resources (NCRR), a KL2 career development award (RR024990) and UCSD start‐up funding to DEHThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Growth inhibition of breast cancer by two flexible heteroarotinoid enantiomers
Authors: Ginn Emily , Baek Jenny , Zou Hongye , Fallatah Maryam M.J. , Cayton Erica , Liu Shengquan , Louie Maggie 
Published: June 2021

Flexible heteroarotinoid (flex‐hets) are compounds that show promising anti‐cancerous activities. More specifically, SHetA2, a first generation Flex‐Het, has been shown to block the growth of cervical, head and neck, kidney, lung, and ovarian cancers, and most recently, prostate and breast cancers. A second generation of compounds was developed to improve SHetA2's high lipophilicity, limited selectivity, low bioavailability, and its complicated synthesis. Results from our recent study suggest that one of these analogs, 1‐(1‐(naphthalen‐1‐yl)ethyl)‐3‐(4‐nitrophenyl)thiourea (SL‐1‐09) exhibits anti‐cancer activities against both ERα+ and ERα‐breast cancer cells at micromolar concentrations. Since SL‐1‐09 is a mixture of enantiomers, R (SL‐1‐29) and S (SL‐1‐30), each analog was synthesized and purified for further analysis. Our results suggest that SL‐1‐30 shows a greater growth inhibitory effect than SL‐1‐29 against both ERα+ (T47D) and ERα‐(MDA‐MB‐453) breast cancer cells with GI50's of 4.56 ±1.17 μM and 3.03 ±0.86 μM, suggesting that the activity observed in SL‐1‐09 is likely associated with the S enantiomer. Furthermore, preliminary cell cycle analysis suggests the percent of MDA‐MB‐453 cells in S‐phase is reduced by 25% when treated with 5.0 μM SL‐1‐30 for 24 hours and 20% reduced when treated with 5.0 μM SL‐1‐30 for 48 hours. Consistent with this data, both T47D and MDA‐MB‐453 express lower levels of cell cycle proteins and genes—cyclin A, cyclin B1, cyclin D1, cyclin E and CDK2 48 hours after treatment with SL‐1‐09 and SL‐1‐30, with cyclin E and CDK2 showing significant changes as early as 12 hours in cells treated with SL‐1‐30. In summary, these results demonstrate that both SL‐1‐30 and SL‐1‐09 inhibit ERa+ and ERa− breast cancer cell growth, potentially by blocking cell cycle progression; however further studies are necessary to delineate the mechanism of action.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Real Time PCR analysis of Ewing's Sarcoma Cell Lines in Response to Chemotherapeutic Treatment.
Authors: Pitzen Sam , Kruchten Anne 
Published: June 2021

Ewing's Sarcoma is a rare bone and soft tissue cancer typically found in children ages ten to twenty years old. Survival rate is about 70% if caught before metastasis, but drops to 30% if the cancer has metastasized throughout the body. Treatment options for Ewing's Sarcoma include amputation, radiation, and/or chemotherapy. To better understand the effects of chemotherapeutic drugs on cultured Ewing's Sarcoma cell lines, we treated Ewing's cells with etoposide, one of the common drugs of the chemotherapeutic cocktail treatment for Ewing's, and with tamoxifen. We used real‐time PCR to analyze mRNA levels of several targets in cells treated with these various chemotherapies. One gene target is GLI1, which is highly expressed in primary tumors. Another target gene sequence is TP53, which is normally a tumor suppressor. mRNA was collected from cells treated with the various chemotherapies andy multiple targets were analyzed. Here we report the results of real time PCR using Taqman probes. Our results contribute to our understanding of how chemotherapies may effect Ewing's Sarcoma tumors and future cultured cell line research in this area.Support or Funding InformationThis work is funded by the College of Saint Scholastica SSC Summer Research Fellowships.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Investigate Kentucky Hemp‐Induced Modulation of Interleukin‐1 β Secretion in Ovarian Cancer Cells
Authors: Turner Chase , Kenley Colby , Jent Travis , Vu Nam , Hughes Nathan , Sumanasekera Wasana K. 
Published: June 2021

Our laboratory is interested in searching for alternative therapies for ovarian cancer. Based on our preliminary studies, KY grown hemp, which is similar to marijuana yet non‐addictive, demonstrated anti‐cancer effects against ovarian cancer. The objective of this study is to investigate the mechanism behind the hemp‐induced anti‐cancer effects. Inflammation is known to play a role in carcinogenesis and Interleukins are contributing players of the inflammation. We hypothesized that the hemp‐induced modulation of Interleukin‐1 β (IL‐1 β) production may play a role in hemp‐induced anti‐cancer effects. Both marijuana and hemp contain therapeutically valuable components such as Cannabidiol (CBD), Cannabidiolic Acid (CBD‐A) and Tetrahydrocannabinol (THC). However, hemp is more beneficial than marijuana due to its' non‐addictive properties. CBD‐A and CBD‐induced modulation of cytokines have been reported in different cell types. To our knowledge, hemp‐induced modulation of ovarian cancer and responsible mechanisms are yet to be discovered.MethodsKY hemp extracts were made using supercritical CO2 extraction method by KY commonwealth Extracts company. CBD content of KY hemp was analyzed using High Performance Liquid Chromatography (HPLC) by MRX laboratories. In our laboratory, these hemp extracts were tested for its' potential anti‐cancer effects and mechanism of action. A2780 epithelial ovarian cancer cells (OCCs) were seeded in 12–well plates. Following serum starvation, OCCs were treated with either vehicle or different concentrations of hemp extracts for 24 hours. Then, cell supernatants were removed and subject to Bradford protein assay followed by IL‐1 β sandwich ELISA. IL‐1 β released from each treatment group was detected based on manufacturer's protocol. In brief, IL‐1 β standards, cell supernatants from treatment groups, and no hemp negative control were incubated in ELISA plate coated with IL‐1 β antibodies. After a series of steps that included secondary antibodies and a substrate solution, IL‐1 β levels were detected spectrophotometrically. For an experiment, each treatment was done in triplicate and each experiment was repeated a minimum of three times to ensure reproducibility. Hemp‐induced attenuation of IL‐1 β secretion was detected and this attenuation is significant at 500 μg/ml hemp extract treatment, which contains 335 μM CBD and 591 μM CBD‐A. Based on the data, it can be concluded that the hemp extract‐induced attenuation of IL‐1 β secretion might contribute to its protective effects against ovarian cancer.Support or Funding InformationFunding was provided by Sullivan University System via a faculty development grant awarded to Dr. Sumanasekera. Grant number: RG_1_PS_2017_01This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evaluating the Role of the Na+‐H+ Exchanger Isoform 1 (NHE1) in Lysophosphatidic Acid and Urokinase‐type Plasminogen Activator Stimulation of Ovarian Cancer Cells
Authors: Stiglich Ashley A. , Provost Joseph J. , Wallert Mark 
Published: June 2021

Lysophosphatidic Acid (LPA) is a bioactive lipid which stimulates an increase in cell proliferation and migration in a range of normal and cancer lines in part through the activation of both the RhoA/Rock and Ras/ErK signaling pathways. Plasma levels of LPA are elevated in ovarian cancer patients even in the early stages of the disease where it is believed to participate in pathogenesis. In patients with more advanced stages of ovarian cancer, the higher the levels of LPA found in the blood and ascites is a negative prognostic indicator. In ovarian cancer, LPA enhances cell proliferation and promotes cell survival by preventing apoptosis and anoikis. LPA also decreases the sensitivity of the ovarian cancer cells to cisplatin and other chemotherapy drugs. While LPA signaling can directly enhance ovarian cancer progression, it can also act by stimulating the production and release of urokinase‐type plasminogen activator (uPA). uPA is a serine protease that binds to a plasma membrane receptor stimulating activation of both the RhoA/Rock and Ras/ErK signaling pathways. Studies in non‐small cell lung cancer cells have shown that LPA and uPA activate the Na+‐H+ Exchanger Isoform 1 (NHE1) leading to an increase in cell proliferation and migration. NHE1 is an 815 amino acid transmembrane protein that exchanges 1 extracellular Na+ for 1 intracellular H+ leading to the pH inversion common to cancer cells in a developing tumor. Activation of NHE1 is one of the initial steps in the development of the transformed phenotype of cancer cells. NHE1 can be phosphorylated and activated by Rock and kinases in the Ras/Erk signaling pathway. Recently, increases in NHE1 expression was demonstrated to play an important role in cancer progression in ovarian cancer patients. In this project, we specifically explore the relationship between LPA and uPA signaling to NHE1 in the regulation of cell proliferation, stress fiber formation, and cell migration in three ovarian cancer cell lines, CAOV‐3, SKOV‐3, and OVCAR‐3.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of HSF2 in Human Malignancies
Authors: Takagishi Seesha , Metz Kyle , Wu Yaqi , Alasady Milad , Mendillo Marc 
Published: June 2021

Heat‐Shock Factor 1 (HSF1) is the master transcriptional regulator of the heat shock response (HSR), an ancient survival mechanism that is conserved from yeast to humans. In the HSR, proteotoxic stress activates HSF1 to induce expression of heat shock protein genes, including HSP70 and HSP90, to maintain protein homeostasis. In cancer, HSF1 is highly expressed, and drives a transcriptional program distinct from the HSR. Genes in the HSF1 cancer program are involved in a broad range of cellular activities that include protein folding, but also include cell proliferation, cell adhesion, apoptosis, and energy metabolism. To better understand HSF1 function in cancer cells, we used a multifaceted strategy to identify HSF1‐interacting proteins. We found that Heat‐Shock Factor 2 (HSF2), an HSF1 paralog, was among the strongest HSF1 interacting proteins. However, the role of HSF2 in cancer cells and its influence on HSF1 activity is not well understood. We performed RNA‐Seq and ChIP‐Seq to identify the HSF2 cancer program. We used CRISPR‐Cas9 mediated genome editing to knock out HSF2 and HSF1 in a diverse panel of common human cancer cell lines. We found that HSF2 and HSF1 exhibit distinctive effects on transcription, despite sharing a high degree of genomic co‐occupancy. These transcriptional effects were affected by the type of cancer cell line. Lastly, we characterized the HSF2 cancer phenotype in vitro and in vivo. In vitro, we found that the loss of HSF2 affects cell migration and invasion. In vivo, loss of HSF2 affects tumor formation and growth. Our findings suggest a complex interplay between HSF1 and HSF2 that ultimately impacts malignant progression.Support or Funding InformationNIH/NCI R00 CA175293‐03, Kimmel Scholars Program SKF‐16‐135, Lynn Sage FoundationThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Endothelial‐derived Extracellular Vesicles Induce Proliferation in Glioblastoma Cells
Authors: Zimmerman Zachary R. , Dores Michael Robert 
Published: June 2021

Extracellular vesicles (ECVs) are critical biomarkers for disease and mediate intracellular signaling within the tumor microenvironment. ECVs from tumors enhance cell proliferation and migration in endothelial cells. However, the role of endothelial ECVs in regulating tumor cell proliferation is not well understood. We have investigated the role ECVs play in the communication between glioblastoma cancers and the human endothelium. To determine the effect of ECVs on the communication between glioblastoma multiforme cancer and endothelial cells, ECVs from both cell types were isolated. These ECVs were determined to carry the proteins CD63 and ALIX, known exosome biomarkers and proteins found in extracellular vesicles originating from the multivesicular body. Glioblastoma cells treated with ALIX‐positive exosomes collected from endothelial cells demonstrate increased migration over untreated cells. Signaling through G protein‐coupled receptors modulates ECV content in endothelial exosomes and increases the amount of ALIX‐positive ECVs released. We are investigating whether changing ECV content through GPCR signaling increases their ability to induce proliferation in glioblastoma cells.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.