bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of TRPP2 binding partners in mice tissues
Authors: Ng Courtney , Yu Yong 
Published: November 2023

Mutations in transient receptor potential polycystin‐2 (TRPP2 or PC2) and its related protein, polycystin‐1 (PKD1 or PC1) result in autosomal dominant polycystic kidney disease (ADPKD), one of the most common human genetic diseases. TRPP2 and PKD1 form a receptor/ion channel complex, which has been proposed to function in coupling extracellular stimuli to intracellular Ca2+ signals. The study of these proteins is essential to understanding the characteristics of this channelopathy and developing specific treatments. Our focus has been working with TRPP2 in order to gain insight on the molecular mechanisms of its structural assembly, and channel function and regulation. TRP channels are localized in a variety of tissues and perform specific roles in sensory physiology. Specifically, TRPP2 is known to express in the human heart, brain, kidney, lung, testis, and ovary. It is also known to have direct binding to a number of proteins, many of which are involved in cytoskeletal association, trafficking and ion channel modulation. The intracellular C‐terminus of TRPP2 has been shown to be involved in the association of some of these binding partners. We believe that the C‐terminus of TRPP2 has major implications in diversifying its role in different tissue types. The approach taken here was to identify novel TRPP2 C‐terminal binding partners present in different tissue types. We purified the C‐terminus of human TRPP2 by expressing it in E. coli, and used it to pull‐down these possible binding partners in several mice tissues, including the brain, kidneys, heart, lungs, and liver. Potential partners were isolated and mass spectrometry was used to identify these proteins for further analysis and functional study. The results of this study will shed light on the specific physiological function of TRPP2 in particular organs.Support or Funding InformationThis research is funded NIH Grant DK102092.
Hydrogen Sulfide Oxidation by Myoglobin
Authors: Bostelaar Trever Michael 
Published: November 2023

Enzymes in the sulfur network generate the signaling molecule, hydrogen sulfide (H2S), from amino acids cysteine and homocysteine. Since it is toxic at elevated concentrations, cells are equipped to clear H2S. A canonical sulfide oxidation pathway operates in mitochondria, converting H2S to thiosulfate and sulfate. We have recently discovered the ability of ferric hemoglobin to oxidize sulfide to thoisulfate and iron‐bound hydropolysulfides. In this study, we report that myoglobin exhibits a similar capacity for sulfide oxidation. We have trapped and characterized iron‐bound sulfur intermediates using cryo‐mass spectrometry and X‐ray absorption spectroscopy. Further support for the postulated intermediates in the chemically challenging conversion of H2S to thiosulfate and iron‐bound catenated sulfur products is provided by EPR and resonance Raman spectroscopy.Support or Funding InformationThis work was supported in part by grants from the National Institutes of Health (GM112455 to R.B. and DK068139 to T.L.S.) and the American Heart Association (14POST18760003 to P.K.Y.). We thank Dr. Markus Ruetz for help with the EPR studies, Andrew Hunt for help with the resonance Raman studies, and Dr. David Ballou for his assistance with the O2 consumption assays.
Induction of apoptosis in carcinoma cells: Perhaps it is right way to kill the metastatic process
Authors: Basu Subhash , Basu Manju , Ma Rui , Moskal Joseph R 
Published: November 2023

The normal cells in their life cycle die through apoptosis (programmed cell death or degradation of genomic DNA). On the other hand, metastatic cancer cells die mostly by Necrosis where toxic gene products come out through the holes on the cancer cell surface to induce necrotic death‐effect on normal cells. In cancer cells, the following chemical could inhibit blocked steps for apoptosis: cis‐platin (commonly used drug for testicular cancer treatment), Betulinic acid (a naturally occurring triterpene), L‐PPMP, D‐PDMP (inhibitor of GlcT [ceramide: glucosyltransferase]), Tamoxifen (commonly used drug for breast cancer treatment), and Melphalan (Golgi bodies scrambler). When three clones of metastatic breast cancer cells (SKBR‐3, MCF‐7 and MDA468) were treated with these apoptotic reagents separately, they induced apoptosis [activated Caspases (‐3, ‐8, or ‐9)] and down‐regulated the genes for cancer markers like sialo‐LeX (determined by DNA‐microarray studies). Targeted application of apoptotic chemicals to kill the cancer cells would be an ideal way for breast cancer therapy.Support or Funding InformationSupported by NIH‐CA‐14764 and NIH‐NS‐18005
Genetic Characterization of Programmed Cell Death in Aneuploid Yeast Cells
Authors: Sanborn Matthew , Timmel Victoria , Barrios Jessie , McBride Trevor , Pinches Robert Seth , Austriaco Nicanor 
Published: November 2023

Aneuploidy is the genetic state of a cell that has a chromosomal number that is not an exact multiple of the haploid complement. It is a leading cause of spontaneous abortions and of mental retardation in humans, and is a characteristic defect in cancer. Yeast cells that are aneuploid manifest a diversity of phenotypes including cell cycle defects, genomic instability, protein imbalance, chaperone stress and proteotoxicity. We are investigating the links between aneuploidy on the budding yeast, Saccharomyces cerevisiae and programmed cell death. We have shown that aneuploid yeast cells are more sensitive to ethanol‐induced cell death. In addition, we have also shown that caloric restriction protects aneuploid yeast from programmed cell death.Support or Funding InformationIn addition to funding from the RI‐INBRE Program, our laboratory is supported by grant NIGMS R15 GM110578, awarded to N. Austriaco.
The Effect of Compound L19 on Human Colorectal Cells (DLD‐1)
Authors: Mohammadhosseinpour Sepideh , Clack Beatrice A. 
Published: November 2023

Colorectal cancer (CRC) is the third most common type of cancer in the world and the second leading cause of human deaths due to cancer (Siegel, et al. 2014). Rumex crispus extracts, a source of anticancer compounds, have demonstrated the induction of apoptosis in colorectal cancer cells (DLD‐1). Bhandari (2015) screened water soluble compounds from roots and leaves identifying those that induced apoptosis in DLD‐1 cells. L19 is a compound that will be studied in more detail in this project. In this research, specific genes involved in apoptosis induction as a result of treatment of the DLD‐1 cells with L19 are identified using methods already utilized by Bhandari (2015) with addition of qRT‐PCR analysis. Accelerated solvent extraction (ASE) followed by high performance liquid chromatography (HPLC) was utilized to obtain larger amounts of L19. GC‐Mass spectrometry was used to identify the isolated compound. Apoptosis was measured by detecting the levels of caspase 3 and 7 using the APO‐one assay (Promega, Corp.) after treating the DLD‐1 cells with varying concentrations of L19. Furthermore, the mechanism of apoptosis has been determined using qRT‐PCR and gene specific primers for caspase 8,10 (extrinsic pathway), 9 (instrinsic), 4, 5 and 12 (ER stress).Support or Funding InformationOffice of Research and Sponsored Projects, SFASU, Ed and Gwen Cole
Understanding the role of BCL‐2 Proteins in Hyperglycemia‐induced Apoptosis in Cardiomyocytes
Authors: Miller Stefanie , Del Gaizo Moore Victoria 
Published: November 2023

Chronic high glucose, or hyperglycemia, is a common marker for type‐2 diabetes and has been linked to diabetic cardiomyopathy. Specifically, when a secondary stressor is introduced, such as oxidative stress, increased cardiomyocyte apoptosis results leading to a larger area of dysfunction. Apoptosis is controlled by the BCL‐2 family of proteins, which consists of both pro‐apoptotic and anti‐apoptotic proteins that either facilitate or restrict cell death, respectively. BCL‐2 proteins interact at the mitochondria and changes to the balance of anti‐and pro‐apoptotic proteins can induce apoptosis. However, there are times during certain types of cellular stress where apoptosis does not occur but rather anti‐apoptotic proteins are sensitized, known as mitochondrial priming. Priming decreases the anti‐apoptotic reserve of the cell, thereby making it easier to antagonize apoptosis upon any further stress. We hypothesize that heart cells chronically exposed to high glucose levels leads to mitochondrial priming. Using a rat cardiomyocyte line, H9C2 cells, a hyperglycemia model was established where cells are cultured in low glucose media and then switched to high glucose media for various amounts of time. To investigate priming, a secondary stressor of diluted H2O2 is then used to induce oxidative stress. The amount of apoptosis in different treatment groups (low glucose, low glucose + H2O2, high glucose, and high glucose + H2O2) was quantified by flow cytometry using Annexin V‐FITC (FL1‐A) and Propidium Iodide (FL3‐A), for early apoptosis and late apoptosis, respectively. Increased levels of apoptosis occurred in cells treated with high glucose media for 72 hours followed by H2O2 (67.8% for FL1‐A and 51.1% for FL3‐A), but not high glucose media alone (9.1% for FL1‐A and 3.0% for FL3‐A). In fact, cells cultured in high glucose media were comparable to those in low glucose with H2O2 (12.2% for FL1‐A and 3.8% for FL3‐A) and without H2O2 (12.9% for FL1‐A and 3.9% for FL3‐A). The levels of apoptosis for the high glucose + H2O2 trial also increased as the time treated with high glucose increased from 48 hours to 72 hours, with values for FL1‐A of 30.6% and 67.8%, respectively. These results demonstrate that high glucose alone does not induce apoptosis, but rather primes cells for apoptosis so when a secondary stressor occurs, cells have an increased propensity to undergo apoptosis.Support or Funding InformationThe authors would like to recognize the following sources of funding: NC Biotech 2013‐IDG‐1019 (VDGM), Elon University Honors Program (SM), Elon U. Undergraduate Research Program (SM and VDGM), and Elon U. FR&D (VDGM).
UV irradiation/cold shock‐induced bubbling death is Ca2+‐dependent
Authors: Tsai Cheng‐Chang , Chen Yan‐Wei , Chang Nan‐Shan 
Published: November 2023

Bubbling cell death is a non‐apoptotic event that each cell generates a single bubble from the nucleus in response to UV irradiation and/or cold shock. UV energy is first absorbed by the nucleus, followed by rapid relocation of cytosolic WWOX, p53, NOS2, TRAF2, and many others to the nucleus to generate a nitric oxide‐containing bubble in the nucleus. The single enlarging bubble merges with cell membrane and then blasts off from the cell to cause cell death. Unlike apoptosis, bubbling‐induced death is irreversible. The death event is caspase‐independent and devoid of DNA fragmentation. Tumor necrosis factor receptor‐associated factor 2 (TRAF2) inhibits bubbling death, while tumor suppressors p53 and WWOX do the opposite. Exosome release occurs during bubbling death, but not in apoptosis. The entire nucleoli may dump into the bubble. In certain cells, damaged nucleoli release the nucleolar contents directly to the bubble. Notably, housekeeping proteins, such as α‐tubulin, are significantly down‐regulated in L929 and COS7 fibroblasts and other cell types. When Ca2+ was depleted from environment, UV‐induced nuclear bubbling was blocked. A likely mechanism is that without Ca2+, executor proteins such as p53, NFκB‐p50, TRAF2 and NOS2 do not translocate to nucleus. The filamentary structure of α‐tubulin is disrupted upon exposure to UV/cold shock. Gene expression profiling in L929 cells revealed that mRNA processing, mRNA metabolic process and RNA splicing are down‐regulated, whereas sensory perception of smell, sensory perception of chemical stimulus and G‐protein coupled receptor protein signaling pathway are up‐regulated. In conclusion, we determined that UV irradiation/cold shock‐induced bubbling death at low temperature is via Ca2+influx for leading to nuclear translocation of executor proteins along with shut down of RNA processing machinery.Support or Funding InformationMinistry of Science and Technology, Taiwan
Stay On Target: Deconvoluting Mixed Redox Messages through Precision Redox Targeting
Authors: Poganik Jesse R. , Parvez Saba , Aye Yimon 
Published: November 2023

The precisely timed and spatially regulated chemical signals are the essence of biochemical redox signaling. However, defining the precise biological impacts of localized signals that engage with specific protein targets under physiologic conditions has proven to be highly challenging. This abstract presents a unique set of proximity‐directed chemical tools that enables powerful interrogation into functional consequences of specific redox events through precision redox targeting in living systems (see, for example, Parvez et al. Nature Protocols 2016 11 2328; Long et al. 2016 Nature Chemical Biology Accepted). With this in vivo‐validated redox‐targeting toolset, we identify bona fide “first responders” sufficient to drive phenotypic responses at organismal level exemplified by zebrafish and C. elegans models. Our unique chemical biology toolset sets the stage for ruling in gain‐of‐function (or dominant loss‐of‐function) redox modifications and relating them to phenotype in an unbiased experiment that is not hampered by functional redundancy.Support or Funding InformationThe NSF CAREER, NIH Director's New Innovator, Beckman Young Investigator, and Sloan Fellowship programs are acknowledged for research support.
CCN5/WISP‐2 activates estrogen receptor‐α in normal and cancerous breast epithelial cells and sensitizes them to hormonal therapy
Authors: Sarkar Sandipto , Ghosh Arnab , Banerjee Snigdha , Maity Gargi , Das Amlan , Gupta Vijayalaxmi , Haque Inamul , Tawfik Ossama , Larson Melissa , Banerjee Sushanta 
Published: November 2023

BackgroundEstrogen Receptor‐α (ERα)regulation is a complex process and thus it is still unclear why ER‐α islacking in some aggressive breast cancer patients and develops resistance to hormone therapy. It is uncertain if this scenario be reversible by specific genetic manipulation. CCN5 is a member of the CCN (Cyr61, CTGF and NOV) family of matricellular proteins and is an anti‐invasive element in breast cancer progression. CCN5 suppresses the functions of many genes that are linked to various pathological outcomes and aggressive behavior of breast cancer (BC) cells. However, the mechanisms by which CCN5 prevents aggressive phenotypes of breast cancer cells are not entirely clear. CCN5 protein is exclusively expressed in estrogen receptor‐α(ER‐α) positive breast cancer cells, which are weakly invasive/aggressive and rarely metastasize depending on the functional status of ER‐α.ResultsUnique molecular relations between CCN5 and ER‐α has been tentatively established and have been identified as the components of the same signaling pathway that coordinate some metabolic signals associated with the proliferation of ER‐α‐positive breast cancer cells as well as delaying the disease progression from a non‐invasive to an invasivestate. Given the importance of this connection, we sought to determine the role of CCN5 in regulation of ER‐α in different cellular settings and their functional relationship. These studies were carried out in both invitro models and CCN5‐overexpressing transgenic mouse models driven by doxycycline (Dox). Our data shows that CCN5 is sufficient to induce ER‐α expression at the transcription level and functional activity as well as promote and sustain hormonal sensitivity in BC cells. Mechanistically, we have shown that CCN5 upregulates the expression of ER‐alpha through downregulation of AKT activity leading to stabilization and nuclear localization of Foxo3a. Moreover, induction of CCN5/ER‐αsignaling in normal breast epithelial cells in transgenic mice promotes hyperplasia but no carcinogenic growth, suggesting CCN5/ER‐α signaling is a required component to maintain the non‐aggressive niche and hormone sensitivity as well as deferment of the aggressive process.ConclusionCCN5 protein promotes ER‐α activity in both normal and cancer cells and enables cancer cells to respond to anti‐estrogen treatment. CCN5 is a positive regulator of ER‐α in both normal and cancer breast cells.SignificanceCCN5 is required to activate ER‐α. Therefore, resistance to hormonal therapy could be overcome by activation of CCN5 signaling.Support or Funding InformationVA Merit Grant
Attenuation of γ‐secretase mediated activation of Notch signaling induces autophagic cell death in triple negative breast cancer cells
Authors: Das Amlan , Ganguli Arnab , Narayanam Maruthi Kumar , Mukherjee Pritha , Basu Biswarup , Chatterjee Urmi , Banerjee Sushanta , Karmakar Parimal , Kumar Dalip , Chakrabarti Gopal 
Published: November 2023

Notch signaling is reported to be deregulated in several malignancies including breast. γ‐secretase, a multi‐subunit enzyme complex, catalyzes the cleavage of Notch receptor to release the Notch intracellular domain (NICD) in the cytosol, which then translocates into nucleus to activate the downstream signaling. Hence pharmacological inhibition of γ‐secretase might lead to the subsequent inhibition of Notch signaling in cancer cells.In search of novel γ‐secretase inhibitors (GSIs), we screened a series of triazole‐based compounds, for their potential to bind γ‐secretase and we observed that 3‐(3′4′,5′‐Trimethoxyphenyl)‐5‐(N‐methyl‐3′‐indolyl)‐1,2,4‐triazole compound (also known as NMK‐T‐057) can bind to γ‐secretase complex. Very interestingly, NMK‐T‐057 was found to inhibit proleiferation, colony forming ability, motility in various triple negative breast cancer cells such as MDA‐MB‐231, MDA‐MB‐468, 4T1 and also MCF‐7, with a negligible cytotoxicity against non‐cancerous MCF‐10A, PBMC and WI38. Moreover NMK‐T‐057 treatment resulted in the reprogramming of the EMT process in TNBCs, accompanied by the drastic reduction of CD44high/ CD24low population. NMK‐T‐057 also inhibited the spheroid forming ability of TNBCs and reduced the stem cell enriched side population. It was also observed that in NMK‐T‐057 treated cells, Notch‐1 and its downstream targets such as Hes‐1, pAkt, and Cyclin D1 were significantly down regulated. The in silico study revealing the affinity of NMK‐T‐057 towards γ‐secretase, was further validated by fluorescence based γ‐secretase activity assay, which confirmed the inhibition of γ‐secretase activity in NMK‐T‐057 treated TNBC cells.Autophagy plays a dual role in cancer depending on its role in the survival or death of cancer cells. The relationship between autophagy and Notch signaling is not clear and only a few reports have come across in highly reputed journals in last one year. We observed that NMK‐T‐057 induces significant autophagic responses in TNBCs and administration of the autophagy inhibitor 3‐MA, attenuates NMK‐T‐057 induced cell death. Similar results were observed on the treatment of TNBC cells with commercially available γ‐secretase inhibitor DAPT. Thus we might conclude that NMK‐T‐057 could be potential drug candidate against breast cancer, which can trigger autophagy mediated cell death in breast cancer cells by inhibiting the γ‐secretase – mediated activation of Notch‐signaling.Support or Funding InformationThis work is supported by Dr. D. S. Korhari Postdoctoral Fellowship Scheme, UGC, Govt. of India.
SRC Regulates Proliferation in ER+ Breast Cancer Cells by Stabilizing MYC mRNA
Authors: Abdullah Christopher , Korkaya Hasan , Courtneidge Sara A 
Published: November 2023

Estrogen (E2) stimulation promotes proliferation in estrogen receptor‐positive breast cancer (ER+ BC), which accounts for 60–75% of all breast cancer. E2 signals through the ER via three pathways: the classical genomic pathway involving transcriptional activation of the ER, crosstalk between ER and other transcription factors, and non‐genomic signaling cascades, which do not involve the transcriptional activity of the ER. The non‐genomic pathway likely involves SRC family kinases (SFKs) and has been implicated in cell cycle downstream of the ER. While not typically amplified or mutationally cancer, SRC is often highly active in most breast cancers.To test whether SRC is required for cell cycle progression in ER+ breast cancer cells, we quiesced ER+ MCF7 or ZR75‐1 cells prior to stimulation with E2 and utilized either a selective SFK inhibitor (SU11333) or shRNA‐mediated knockdown. Inhibition of SFK activity blocked progression from G1 to S phase and proliferation in 2D and 3D assays. We additionally tested the clinically available SRC inhibitor, saracatinib, in an orthotopic xenograft mouse model of ER+ BC and observed decreased tumor growth upon treatment with saracatinib.As MYC is a known target of both SRC and ER and also is known to drive cell cycle progression, we tested whether MYC could be mediating SRC's proliferative effects in this model. Using MYC‐targeting shRNA constructs, we showed that MYC is required for E2‐dependent cell cycle progression and proliferation. Additionally, E2 stimulates MYC mRNA expression dependent upon SRC activity. We also showed that MYC mRNA expression is mediated primarily through SRC‐dependent stabilization of the transcript. We next aimed to understand the mechanism by which non‐genomic ER‐SRC signaling drives proliferation in ER+ BC cells. A known stabilizer of MYC mRNA, the RNA‐binding protein IMP1, has a truncated form (ΔN‐IMP1) expressed in ER+ BC cells. Our data suggests that ΔN‐IMP1 is required for the SRC‐mediated stabilization of MYC mRNA and proliferation. Taken together, these data suggest that SRC mediates cell cycle progression and proliferation through a non‐genomic E2 signaling pathway by stabilizing MYC mRNA.Support or Funding InformationNIH 1F31CA180740 to CANIH 1R21CA177382 to SAC
USING A CRISPR/CAS9 KNOCKOUT TO EVALUATE THE ROLE OF FURIN IN THE INTOXICATION PATHWAY OF PSEUDOMONAS EXOTOXIN A
Authors: Sanford Jack , Zhu Yuyi , Weldon John 
Published: November 2023

Derivatives of Pseudomonas exotoxin A (PE) known as recombinant immunotoxins (RITs) are chimeric proteins being investigated as potential cancer therapies to target and kill cancer cells. RITs gain access to target cells via receptor mediated endocytosis, and traffic through the trans‐Golgi network to the endoplasmic reticulum. From there, RITs gain access to the cytoplasm where they inhibit translation elongation factor 2 (EF2) and halt protein synthesis. The subtilisin/kexin‐like proprotein convertase furin (PCSK3) has been implicated in the PE trafficking pathway as a proteolytic activator of the toxin, but circumstantial evidence suggests that furin may assist intracellular trafficking or have a combination of both cleavage and trafficking functions.This project aims to use the CRISPR/Cas9 methodology to create a furin‐deficient cell line in order to explore the role of furin in the intoxication pathway of RITs. Thus far, a targeting guide RNA has been used to direct the Cas9 endonuclease to the furin gene in HEK293 cells. The resulting cells were clonally expanded and evaluated for knockout mutations resulting from repair of the Cas9 cleavage events. Several HEK293 subclones without furin expression were identified. Initial RIT‐based cytotoxicity assays have shown a difference in susceptibility between the furin knockout and wild type HEK293 cell lines. We plan to introduce furin variants with altered function into the furin‐deficient cell line to quantifiably characterize the role of furin in the RIT intoxication pathway.Support or Funding InformationI would like to thank the Fisher College of Science and Mathematics (FCSM) as well as the Towson Office of Undergraduate Research (TOUR) for their financial support of this project. I would also like to thank all of the members of Dr. Weldon's lab for their help with this research.