bluuebunny/crossref_metadata_2025_split huggingface dataset:
Nuclear Trafficking of ABCB1‐Daunorubicin Vesicles Initiated by Sphingomyelinase Reverts Multidrug Resistance in Chinese Hamster Fibroblasts
Authors: Lee Wing‐Kee , Kolesnick Richard N 
Published: November 2023

Effective cancer chemotherapy treatment is hampered by multidrug resistance (MDR), defined by upregulation of ABCB1, which effluxes drugs. Intracellular ABCB1 also sequesters anthracycline drugs in acidic vesicles preventing nuclear targeting. Reports indicate sphingolipids affect vesicular trafficking. We hypothesized sphingomyelinase (SMase) activity might mobilize ABCB1/drug vesicles to revert MDR. Using drug‐resistant ADX fibroblasts expressing ABCB1‐EGFP, time‐lapse confocal microscopy revealed daunorubicin (DNR) sequestration in ABCB1‐EGFP‐containing vesicles within 5min. In ADX/ABCB1‐EGFP loaded with DNR (20μM,2h) followed by drug washout, exogenous bacterial (b)SMase, which hydrolyzes plasma membrane sphingomyelin and initiates sphingolipid signaling, induced rapid ABCB1‐EGFP/DNR vesicle movement to the nucleus, where DNR was deposited, while ABCB1‐EGFP remained in nuclear membranes. Immunoblots of isolated nuclei and nuclear envelope membranes purified by sucrose density gradient confirmed these observations. In DNR‐exposed ADX cells, bSMase‐induced ABCB1 translocation to the nucleus peaked at 1h, and was detectable at 4h but not at 8h, suggesting transient incorporation of ABCB1 into the nuclear membrane. No nuclear ABCB1 was observed in drug sensitive DC3F cells loaded with an isotoxic DNR dose (0.2μM, 2h), indicating ABCB1 nuclear translocation by bSMase is MDR specific. Cell viability studies by MTT assay demonstrate augmented DNR toxicity in the presence of bSMase and can be enhanced by co‐incubation with the ABCB1 inhibitor, PSC833/valspodar (1μM), to prevent cellular drug efflux, in an additive manner. Intriguingly, in the postnuclear supernatant of DNR‐treated ADX cells, ABCB1 primarily sediments at 8000g, but shifts to 18000g at 4h after bSMase suggesting altered vesicular density or ABCB1 transfer to a distinct vesicular pool. In conclusion, SMase activity mobilizes ABCB1/DNR vesicles to the nucleus where DNR is deposited and reverses MDR.Support or Funding InformationThe work was supported by the Max Kade Foundation and Intramural Research Program at Witten/Herdecke University (IFF2016‐20) (W.K.L).
Optimization of Schwann Cell Differentiation from Porcine Stem Cells
Authors: Schomberg Dominic , Shanmuganayagam Dhanansayan 
Published: November 2023

Swine as a translational animal model of cancer is becoming increasingly recognized for its value in closing the gap between basic research and the development of clinically effective therapies. While rodent models have helped elucidate the complexities of many mechanistic pathways of cancer, they often have physiological and genetic differences that greatly limit the translatability of results to humans. The recognition of the value of swine models has also created a need for swine‐based in vitro cell culture platforms for mechanistic studies and drug screening. Such platforms are particularly sought after for tackling Schwannomas (Schwann cell based tumors) that are associated with diseases like neurofibromatosis 1 (NF1), a genetic disease caused by a mutation of the neurofibromin 1 (nf1) gene. These tumors are often benign, however, a small portion of them become malignant, degenerating into a form of cancer called neurofibrosarcoma. Patients with NF1 often develop neurofibromas from Schwann cells that can become malignant peripheral nerve sheath tumors, a tumor type that carries poor prognosis. Schwann cells, like most cells of a neuronal lineage, are challenging to culture in vitro. This has slowed progress of understanding the mechanisms responsible for these types of cancers. We therefore aim to develop an in vitro culture platform using swine cells to overcome these obstacles. This necessitated the optimization of culturing conditions to differentiate porcine stem cells into Schwann cells and then develop tumorigenic stem cells by the manipulation of the nf1 gene.We isolated adipose‐derived mesenchymal stem cells and testis‐derived spermatogonial stem cells for use in our experiments. Culturing parameters manipulated include temperature, matrix composition, growth factor levels, feeder cell status, and passaging conditions. In the subsequent phase, different stem cell transfection methods were studied with the goal of delivering CRISPR/Cas9 components targeting the nf1 gene. Techniques compared include lipofection using Lipofectamine 2000 and 3000 and electroporation using the Gene Pulser II or Neon Transfection System. Our findings will help researchers establish a more robust in vitro model for culturing Schwann cells from swine stem cells to study cancer development.Support or Funding InformationThe research was supported by the Biomedical & Genomic Research Group Discretionary Fund (University of Wisconsin‐Madison), NF Network, NF Central Plains, NF Michigan, NF Northeast, and Washington State NF Family.
African Americans with Pancreatic Ductal Adenocarcinoma Exhibit Gender Differences in Kaiso Expression
Authors: Mukherjee Angana , Jones Jacqueline , Yates Clayton 
Published: November 2023

Kaiso, a bi‐modal transcription factor, regulates gene expression and is elevated in breast, prostate, and colon cancers. Depletion of Kaiso in other cancer types leads to a reduction in markers for the epithelial– mesenchymal transition (EMT), however, its clinical implications in pancreatic ductal adenocarcinoma (PDCA) have not been widely explored. PDCA is rarely detected at an early stage but is characterized by rapid progression and invasiveness. Herein, we report the significance of the subcellular localization of Kaiso in PDCAs from African Americans. In the overall patient population, we observed a higher expression of cytoplasmic Kaiso in high‐grade pancreatic tumors compared to low‐grade pancreatic tumors (P = 0.0435). Kaiso expression was higher in the cytoplasm of invasive and metastatic pancreatic cancers (P < 0.0001). In males, cytoplasmic and nuclear expression of Kaiso correlated with cancer grade and lymph node positivity (P = 0.0013; P = 0.0112 and P = 0.0177, respectively). In male and female patients, cytoplasmic Kaiso expression correlated with invasiveness (P = 0.0101). Further, analysis of the largest PDCA dataset available on ONCOMINE shows that with an increase in Kaiso expression, there is an overall increase in Zeb1, which is the inverse for E‐cadherin. Hence, these findings suggest an oncogenic role for Kaiso in the progression of PDCAs, involving the EMT markers, E‐cadherin and Zeb1.Support or Funding InformationThis work was supported by the Department of Defense Prostate Cancer Research Program (PC120913) [CY], G12 RR03059‐21A1 (NIH/RCMI) [CY], (NIH/NCI) 1 R21 CA188799‐01 [CY] and U54 CA118623 (NIH/NCI) [CY], a pilot project on Pancreatic SPORE 2 P50 CA101955‐11 [CY], Statistical analysis was performed by Tuskegee University Statistical Core.
Aspirin attenuates melanoma tumor growth via PGF2α‐SOX‐2 dependent pathways
Authors: Thyagarajan‐Sahu Anita , Sahu Ravi P 
Published: November 2023

IntroductionExposure to pro‐oxidative stressors including cancer therapies via its ability to generate reactive oxygen species (ROS) produce potent oxidized lipid mediator, platelet‐activating factor (PAF) and PAF‐like agonists. We have shown that these PAF‐agonists significantly: 1) augmented the growth of subcutaneously implanted murine B16F10 melanoma tumors in a PAF‐receptor (PAF‐R) dependent manner; 2) impeded the efficacy of cancer chemotherapeutic agents and radiation therapy in murine melanoma models and; 3) detected in the perfusates and tumors collected from chemotherapy and radiation therapy‐treated melanoma and non‐melanoma cancer patients. As melanoma tumors often acquire resistance to the known chemotherapy, radiation therapy and even novel targeted therapeutic regimens, it is necessary to investigate an alternative approach for its intervention. Aspirin (ASN) has classically been used as an anti‐inflammatory drug and also possesses anti‐cancer activity in‐part via its ability to inhibit enzyme cyclooxygenases. While anti‐carcinogenic activity of ASN has been shown in other cancers, little is known about its effects in melanoma.ObjectivesThe current studies were sought to investigate the mechanism and test the hypothesis if ASN‐mediated anti‐melanoma activity is via inhibition of PAF‐R activation.MethodologyCell proliferation and survival assay, apoptosis by caspase 3/7 luminescence assay, immunoblotting, gene array, quantitative real time PCR and flow cytometry studies were performed. These assays were conducted in stably PAF‐R‐expressing (B16‐PAFR) and deficient melanoma (B16‐MSCV) cells and C57BL/6‐wild‐type (WT) and PAF‐R knockout (PAFR−/−) mice.Results and DiscussionWe demonstrate that ASN inhibits the survival of B16‐PAFR and B16‐MSCV cells at similar rates in a time and dose dependent manner via inducing apoptosis as measured increased caspase 3/7 activity. These ASN‐mediated effects were blocked by PGF2α addition but not by PGE2. Importantly, ASN‐mediated significant suppression of murine B16‐PAFR and B16‐MSCV melanoma tumor growth was independent to both cellular and stromal PAF‐R expression. Moreover, gene array analysis of melanoma tumors revealed an inhibition of SOX‐2 oncogene by ASN. Furthermore, PGF2α add back upregulated SOX‐2 and rescued melanoma cells from ASN‐mediated inhibition of cell survival and induction of apoptosis.ConclusionASN targets PGF2α and downstream SOX‐2 to inhibit melanoma tumor growth in a PAF‐R independent pathway and provide the rationale for its implication as a novel melanoma therapy.Support or Funding InformationNIH K22 ES023850 and WSU‐RIF 190002
Expression of matricellular protein CCN1 in the tumor stroma is required for melanoma metastasis
Authors: Leask Andrew , Postovit Lynne , Quensel Katherine , Hutchenreuther James 
Published: November 2023

Metastatic melanoma has an extremely poor prognosis. BRAF inhibitors are used to treat a subset of patients but eventually patients acquire resistance to this therapy. An increasing body of evidence suggests that an activated tumor stroma is responsible for the acquisition of resistance to chemotherapy. Herein, we find that he secreted matricellular protein cyr61(CCN1) is overexpressed in human melanoma in a fashion that correlates negatively with clinical outcome, positively with the extent of tumor stroma development and not at all with BRAF mutational status. We have used a fibroblast‐specific type I collagen promoter‐driven cre recombinase that is tamoxifen dependent to generate C57BL6 mice deleted or not for CCN1 in fibroblasts. CCN1‐deficicent mice display progressive reduction in skin thickness. When injected subcutaneously, B16(F10) cells show reduced metastasis to the lung. Tumor growth is unaffected. This phenotype is linked with reduction in expression of collagen‐modifying enzymes LOX, P4H and PLOD2 as well as a decrease in insoluble collagen protein expression as visualized by a hydroxyproline assay and by trichrome staining. Type I collagen mRNA expression is unaffected.. Thus, CCN1 is required for proper induction of a tumor stroma and hence for metastasis and thus should be further evaluated as a possible therapeutic target for BRAF inhibitor‐resistant melanoma.Support or Funding InformationCIHR, Canadian Cancer Society
Enlarged endosomes and altered transferrin recycling in breast cancer cells
Authors: Tubbesing Kate Elizabeth , Malhotra Aditi , Rudkouskaya Alena , Barroso Margarida 
Published: November 2023

Understanding how the endocytic pathway is regulated in different types of cancer cells remains a critical biological question with far reaching implications in the development of receptor‐targeted cancer therapies. To address this problem, we have characterized the regulation and organization of the endocytic pathway in triple‐negative breast cancer cells (MDA‐MB‐231), luminal A breast cancer cells (T47D) and non‐cancerous breast epithelial cells (MCF10A). We have focused this study on the well‐known endocytic pathway of transferrin (Tf) and the Rab‐GTPases, which are known regulators of the early, late and recycling endocytic compartments. Super‐resolution microscopy and quantitative confocal morphological analysis have been used to characterize the enlarged Tf and early endosome antigen‐1 (EEA1) positive endosomes in breast cancer cell lines. These enlarged Tf and EEA1 positive endosomes are also observed in human breast cancer tumor xenografts. Our Tf recycling assay shows that T47D cells retain significantly more Tf, compared to MDAMB231 or MCF10A cells. However, the observed Tf recycling behavior in different cancer cells does not correlate with the protein expression levels of transferrin receptor or different endogenous Rab‐GTPases evaluated. Interestingly, the colocalization of EEA1 with a panel of rabs differs between cell lines. This data indicates that differences observed in Tf recycling and endosomal morphology in different cancer cell lines may be dependent on the organization of the Rab‐GTPases, which alters the regulation of the endocytic pathway. Currently, we are performing quantitative live‐cell airyscan microscopy experiments to address the link between the altered endosomal morphology, receptor trafficking and localization of over‐expressed Rab‐GTPases. This research is essential to the optimization of receptor‐targeted anti‐cancer therapies, as the regulation of the endocytic pathway determines the retention, release and ultimate efficiency of the anti‐cancer therapy.Support or Funding InformationAlbany Medical College internal funds
MiR‐105/93‐3p promotes metastasis, chemoresistance and stemness in TNBC and circulating miR‐105/93‐3p act as predictive biomarker for TNBC chemoresistance
Authors: Li Hao‐Yi , Lu Pei‐Jung 
Published: November 2023

Triple negative breast cancer (TNBC) is defined as lack of ER, PR and HER2 receptor and accounts for 15–20% of all breast cancers. The treatment for TNBC is a big challenge due to lack of hormone and target therapy and neoadjuvant chemotherapy only can achieve 20%–30% pathological complete response in TNBC patients. In the current study, we identified four highly expressing oncomiRs, miR‐301b, miR‐181a‐3p, miR‐105 and miR‐93‐3p that were significantly associated with poor survival in TNBC patients. To explore the oncogenic roles of four oncomiRs described above, these miRNAs were ectopically expressed in MB‐231/HCC70 cells and silenced in HCC1937/BT‐549 cells, respectively, for examining proliferation, metastasis, chemoresistance and stemness. MiR‐105/93‐3p overexpressing cells showed Cisplatin resistance and high metastatic ability whereas silence of miR‐105/93‐3p can decrease Cisplatin resistance, metastatic ability and stemness. Bioinformatics analysis showed that Wnt/b‐catenin is one of the most potential miR‐105/93‐3p downstream pathways. Ectopic overexpression of miR‐105 or miR‐93‐3p actives Wnt/b‐catenin signaling by TOP/FOP reporter assay. Bioinformatics, RNA‐IP and immunoblotting showed that SFRP1 is directly targeted and down‐regulated by miR‐105 and miR‐93‐3p. In addition, restoration of SFRP1 can overcome miR‐105/93‐3p mediated cisplatin resistance. High expression of miR‐105/93‐3p and low expression of SFRP1 strongly associated with TNBC poor survival even after chemotherapy treatment. The expression of miR‐105 and miR‐93‐3p inversely correlate with SFPR1 protein level in TNBC N‐T pair. Notably, circulating miR‐105 and miR‐93‐3p significantly higher expression in TNBC than non‐TNBC patients' serum and normal serum. Collectively, our data conducted that miR‐105/93‐3p activate Wnt/b‐catenin signaling through down‐regulate SFRP1 to promote TNBC poor survival and circulating miR‐105/93‐3p may act as predictive biomarker for TNBC poor survival.Support or Funding InformationMOST‐105‐2320‐B‐006‐054‐
Quercetin inhibition of TNFα‐induced CCL28 release from human triple negative breast cancer (MDA‐MB‐468) cells
Authors: Buchanan Timonia F , Bauer David , Soliman Karam F 
Published: November 2023

Quercetin is a natural chemopreventative flavonoid found in fruits, vegetables, and herbs, which exerts antitumor, antiinflammatory, and antioxidant effects. A hallmark of cancer is the recruitment of regulatory T cells (Tregs), which promotes hypoxia‐driven angiogenesis. In this study, we evaluated the effects of quercetin on the attenuation of the release of tumor necrosis factor‐alpha (TNF‐α)‐induced tumor promoting chemokines in MDA‐MB‐468 triple negative breast cancer cells, with a focus on those associated with Tregs recruitment. Initial studies were performed using antibody microarrays which specifically show that TNF‐α (40ng/mL)‐induced protein expression of CNTF, IGFBP‐1, IL‐5, IL‐6, PARC, TNF‐beta, BTC, CCL28, GITR, GRO, GRO‐alpha, HCC‐4, IL‐8, oncostatin M, sTNF‐RI, VEGR‐D, were downregulated by a sublethal concentration of quercetin (40μM). Furthermore, the obtained data show that quercetin downregulated TNF‐α‐induced CCL28 release by 60% as it was further validated by ELISA. To date, there is little known about the presence of CCL28 in breast cancer. These findings suggest that quercetin can modulate CCL28 release and inhibit the recruitment of Tregs, a pivotal factor in tumor cell growth.Support or Funding InformationThis research was supported by the National Institute for Minority Health and Health Disparities of the National Institutes of Health through Grant Number G12 MD007582 and Grant Number P20 MD006738.
Use of PC‐NSAIDs to treat 2nd degree burn injury in rodent models
Authors: Lichtenberger Lenard M. , Ono Kaori , Phan Tri , Fang Dexing , Poindexter Brian J. , Bick Roger J. , Dial Elizabeth J. 
Published: November 2023

BackgroundThere is a need for drug treatment for 2nd degree burn injury that can be applied quickly to both relieve pain and promote healing. Currently used drugs for chronic pain have substantial side‐effects which limit their usefulness. These include opioids and their addictive potential, gabapentin and its sedation, and coxibs with their risk of cardiovascular failure. Nonsteroidal anti‐inflammatory drugs (NSAIDs) are potent analgesics and provide relief from burn pain, but their use is limited due to side effects related to gastrointestinal (GI) bleeding insusceptible patients.Objective/AimThe goal of this project is to develop rodent model systems to evaluate the analgesic efficacy and side‐effects of a novel treatment for 2nd degree burn injury to alleviate pain and promote healing using a new class of NSAIDs that are complexed with the soy phosphatidylcholine (PC) that can be administered as a sterile topical alone, parenterally or in combination.MethodsEmploying two established methods, for the induction of 2nd degree burn injury in rats to either the dorsal skin or the hindlimb we investigated the effects of the PC‐NSAIDs, Ibuprofen‐PC and Indomethacin‐PC vs the unmodified NSAIDs administered (either subcutaneously (sc) or topically to the affected skin) daily over a 10‐day study at a dose of 20 mg/kg and 2mg/kg respectively. We then assessed pain sensation behaviorally, the depth of skin injury by light and fluorescent microscopy and tissue inflammation biochemically and by non‐invasive IVIS imaging.Results & ConclusionBoth Indomethacin‐PC and Ibuprofen‐PC when sc administered provided moderate hyperalgesia at 3‐ and 5‐days post‐burn injury, which was similar to the unmodified NSAIDs and in some cases more efficacious as analgesic/anti‐inflammatory agents. Topical administration of our test PC‐NSAIDs provided more variable responses using the two rodent model systems, with the most consistent analgesic/anti‐inflammatory effect being recorded with Indomethacin‐PC. Morphological analysis revealed that the depth of skin injury was reduced in rats treated with the PC‐NSAIDs with no evidence of GI bleeding. Supported by a USARMC grant, W81XWH‐14‐2‐0016.Support or Funding InformationSupported by a USARMC grant, W81XWH‐14‐2‐0016.
Analgesic actions of novel derivatives of the active compound, Incarvillateine, from the Chinese herb (Incarvillateine sinesis) in acute pain
Authors: Priebe Alaini , Hunke Megan , Pattabiraman Mahesh , Chandra Surabhi 
Published: November 2023

Commonly used pain medications, opioids and non‐steroidal anti‐inflammatory drugs (NSAIDs) are highly efficacious for treating chronic pain and post‐surgical pain, but are associated with neurological and abdominal side effects. This reduces patient compliance while also leading to abuse and misuse as non‐prescription medication. Thus, there is a high need for formulating alternative pain treatment strategies to investigate drugs with potent action but minimal side effects. Incarvillateine (INCA), derived from the Chinese herb Incarvillea sinensis, has been widely used in traditional medicine for treating rheumatism and pain. Research on antinociceptivity of INCA in chronic and acute pain models indicate that the diaryl cyclobutane dicarboxylic acid core is essential for its medical property. The small number of tested compounds, despite the larger number of possible INCA analogs, is due to the lack of efficient synthetic route to construct the various regio‐ and stereoisomers of the diphenylcyclobutane‐dicarboxylate core. There are reports that though INCA binds to both opioid and adenosine receptors, its primary analgesic action is through the adenosine receptors, thus having minimal side effects on central nervous system. While INCA is a highly effective natural compound, research on synthetic analogs of this compound has received less attention till date. We hypothesized that novel INCA analogs synthesized in our lab show potent antinociceptive action, and their effect is mediated through adenosine receptor action. Compounds were synthesized using innovative cavitand‐mediated photodimerization (CMP) method, which utilizes a macromolecule (gamma‐cyclodextrin) to control the excited state reactivity of photoactive compounds to yield target tetra‐substituted cyclobutanes (dimers). The dimers generated so far show positive response in suppressing formalin‐induced acute pain in mice hind paw while the monomers were ineffective. The antinociceptive effect of these analogs was observed in the inflammatory phase suggesting a primary non‐opioid mechanism for pain reduction. Our further characterization and selection of INCA analogs (with predominant A3AR ‐ adenosine receptor action) will help us to generate a new class of antinociceptives with precise chemical modifications using CMP methodology. The broad goal of this project is to provide a targeted low‐cost approach towards synthesis of novel analgesics with potent action, high efficiency, and minimal aftereffects.Support or Funding InformationAmerican Chemical Society‐Petroleum Research Fund (MP) University of Nebraska‐Kearney Start‐up funds (SC)