bluuebunny/crossref_metadata_2025_split huggingface dataset:
Loss of MET in the Absence of EGFR Signaling Leads to Hepatic Failure in the Resting Liver
Authors: Tsagianni Anastasia , Mars Wendy M. , Bowen William , Haynes Meagan , Paranjpe Shirish , Orr Anne , Michalopoulos George 
Published: November 2023

Our recent study (Paranjpe S. et al, Hepatology 2016 Nov;64(5):1711–1724) showed that in mice, combined systemic elimination of EGFR and MET signaling abolishes liver regeneration after partial hepatectomy, leading to liver decompensation and death at 15–18 days post‐surgery. In the current study, we investigated the role of EGFR and MET in resting livers that were not stimulated by hepatectomy. Exon 16 from MET was removed using Tamoxifen IP injections in MET ff/ff:TamCre+/+ mice, preserving the protein but eliminating efficient signaling through the receptor. Canertinib diet (an irreversible pan‐ERB EGFR inhibitor) was used to achieve a complete block in EGFR signaling. Vehicle corn oil injections and normal chow were used for the control animals. Surprisingly, double inhibited mice were sickened by day 10 with most of them dead by day 14. In the double inhibited group, liver to body weight ratios decreased over time in a statistically significant way at every time point tested (days 2, 7 and 10), whereas ratios with individual treatments were not statistically altered. Further, in the combined MET and EGFR elimination group, MET protein was totally absent while EGFR protein was preserved. As Canertinib diet alone did not lead to the complete loss of MET protein, and as Canertinib completely inhibits EGFR signaling, we interpret these results to mean that EGFR signaling normally stabilizes MET protein through a mechanism involving exon 16 and that the complete loss of MET in the double‐inhibited animals is what ultimately leads to liver failure. These results underscore the fundamental contribution of the combined HGF/MET and EGFR signaling pathways to hepatic proliferation, survival and functionality.Support or Funding InformationMenten Endowment
Role of Na+‐K+‐ATPase in development of alcoholic fatty liver disease
Authors: Matsumoto Hiroshi , Sugimoto Kana , Yang Shipin 
Published: November 2023

Patients with alcoholic fatty liver disease (AFLD) were well‐known to have low blood sodium level. Alcohol intoxication causes low blood sodium level. In the previous study, we obtained that acute and chronic alcohol consumption induced the phenomenon of low blood sodium level in rats. These findings suggested renal failure, but there was no change in renal function in human and rat. Blood sodium level is regulated by Na+‐K+‐ATPase in lots of tissues. Then, we hypothesized that regulation of Na+‐K+‐ATPase may contributes to improve the development of alcoholic organ damage. In the present study, we focused to clarify whether regulation of regulation of Na+‐K+‐ATPase improves alcoholic liver injury in rats or not. Alcoholic fatty liver disease rat model was performed by feeding a Lieber‐DeCalri diet for 4 weeks. Amiloride, lidocaine, ouibain, sanguinarine, or glibenclamide was administered i.p to rats for 2 weeks after feeding an alcohol diet. After 4 weeks feeding, blood biochemical tests were performed. Expression of mRNA and proteins was performed by qPCR and western blotting analysis. EMSA was performed. Histopathological findings showed alcoholic fatty liver changes in the liver of rats fed the alcohol diet On the other hand, administration of inhibitors of Na+‐K+‐ATPase reduced a slight improvement and decreased the NF‐kappaB activity via IRF3‐dependent pathway but increased activation of IRFy‐dependent pathway in the liver, which phenomena spotlighted in the isolated endsome from the liver, leading that Na+‐K+‐ATPase in endosome can be related to activation of innate immune system. These findings suggest that regulation of Na+‐K+‐ATPase may be one of therapies for alcoholic fatty liver disease.Support or Funding InformationThis research was supported in part by JSPS KAKENHI Grant Number 24390175.
Wnt7b and Wnt10a, beta‐catenin independent signaling regulates cholangiocyte proliferation and function during cholestasis
Authors: Kosar Karis , Nejak‐Bowen Kari 
Published: November 2023

Cells found in biliary ducts, are believed to be bipotential cells that are capable of differentiating into both cholangiocytes and hepatocytes, leading researchers to consider them as a sort of stem cells for both the liver and biliary duct. Wnt/beta‐catenin signaling has been implicated as an important factor in hepatoblast atypical ductular reaction and in the liver stem cells of rats and in a variety of liver pathologies, including atypical ductular proliferation during hepatobiliary injury. We have previously shown that liver‐specific b‐catenin knockout (KO) mice exhibited a markedly decreased ductular response after 2 weeks of 0.1% 3,5‐diethoxycarbonyl‐1,4‐dihydro‐collidine (DDC) diet. More recently, we also identified upregulation of specific Wnt proteins in the cholangiocytes during cholestatic liver injury and that mice lacking Wnt secretion from hepatocytes and cholangiocytes (Alb‐cre Wls KO) showed fewer proliferating cholangiocytes and high mortality in response to DDC diet. Therefore, we hypothesize Wnt induces proliferation of cholangiocytes, which may alleviate some of the complications caused by cholangiopathies. Two Wnts of interest for us are Wnt7B and Wnt10A, which we have shown, through in vitro studies, promote proliferation of cholangiocytes in an autocrine manner. These Wnts are specific for cholestasis, as wild‐type (WT) mice show normal expression of Wnt7B and Wnt10A after choline deficient, ethionine‐supplemented diet, which induces hepatocyte injury and ductal hyperplasia unrelated to cholestasis. Proliferation and cell viability were unimpaired in a small cholangiocyte (sm‐cc) cell line after b‐catenin suppression in vitro under normal growth conditions. To confirm these findings, sm‐cc cells were transfected with either Wnt7B or Wnt10A, in combination with either control or b‐catenin siRNA. The absence of b‐catenin did not inhibit the Wnt 7B‐ and 10A‐induced increase in cholangiocyte proliferation. Media from sm‐cc cells transfected with Wnt7B or Wnt10A was also analyzed for cytokine production. Compared to control cells, Wnt7B and Wnt10A induced a significant increase in the expression of cytokines and growth factors such as IL‐1A, IL‐17A, and VEGF, while expression of anti‐inflammatory IL‐2 decreases in the transfected cells. Thus, b‐catenin independent Wnt signaling promotes ductular reaction during cholestasis and may also alter the phenotypic response of proliferating cholangiocytes.Support or Funding InformationNational Institute of Biomedical Imaging and Bioengineering of the National Institute of Health under Award Number T32EB0010216UPP Academic Foundation Award 1R01DK103775
Alcohol abuse promotes breast cancer development and progression via StarD10 phosphorylation
Authors: Floris Andrea , Cossu Carla , Spissu Ylenia , Tomasi Maria Lauda 
Published: November 2023

BackgroundEpidemiological studies demonstrated that ethanol administration promotes breast cancer development and metastasis. Human breast cancer cells or mammary epithelial cells with a high expression of ErbB2 exhibited an enhanced response to ethanol‐stimulated cell invasion in vitro. In addition, ethanol stimulates cell proliferation and triggers different intracellular signaling modulating the phosphorylation status of key proteins as JNK, P38 and ERK. Post‐translational modification by phosphorylation is a common mechanism to regulate the activity of protein, increasing their local negative charge to promote conformational changes or influencing interaction with protein partners. StarD10 is a member of the start protein family well known to be regulated due to Phosphorylation on Serine 284 by Casein Kinase II (CKII); it's a lipid transfer protein with selective binding site to Phosphatidylcholine and Phosphatidylethanolamine. It was found to be co‐expressed with ErbB2 in many breast carcinoma cell lines, suggesting a selective growth advantage for tumor expressing both proteins. The aim of this study was to investigate the role of Phosphorylated StarD10 in breast cancer and its interaction with ErbB2 as consequence of ethanol administration.MethodsThe experiments were performed using MCF7, MDA‐MB231 human carcinoma cell lines treated with ethanol 100mM. Protein and mRNA levels were analyzed by western blotting/Co‐immunoprecipitation (Co‐IP) and RT‐PCR, respectively. The phosphorylation fraction of StarD10 has been detected by protein purification system columns from cell lysate.ResultsWe found that ethanol treatment increases StarD10 and ErbB2 at both protein and mRNA levels in MCF7 cells. Recombinant proteins were used to demonstrate the direct interaction between StarD10 and ErbB2 and we confirmed this finding by Co‐IP. In addition, ethanol treatment resulted in increased StarD10 and ErB2 interaction. We have established that StarD10 and ErbB2 show a reciprocal regulation. Interesting, we found that both overexpression and silencing of Stard10 induce cell growth and migration suggesting an important role in maintaining cellular homeostasis and potential role of StarD10 in induced‐ethanol breast cancer development and/or progression. In addition, we found that ethanol reduces the phosphorylation status of StarD10 via downregulation of CKII, making it more active.ConclusionThe ability of Stard10 to cooperate with ErbB receptors may be independent of its lipid binding function. This novel finding could support the hypothesis of phosphorylation regulates the interaction between ErbB2 and StarD10 and their synergistic action in the neoplastic progression of the breast cancer. This is the first report demonstrating that ethanol can modulate in dynamic manner the phosphorylation status of protein StarD10 in breast cancer cell lines.Support or Funding InformationSupported by NIH grant 5 K01 AA022372‐04(Maria Lauda Tomasi)
A High Fat Diet Augments 4NQO‐induced Oral Squamous Cell Carcinoma in Mice
Authors: Pytynia Matthew , Meyer Alice , Nguyen Andrew , Goral Joanna , Green Jacalyn M , Jham Bruno , Pitstick Lenore , Ciancio Mae J 
Published: November 2023

IntroductionHead and neck squamous cell carcinoma (HNSCC) is the sixth leading cause of cancer worldwide, and is more common in men than women. Since the diagnosis of HNSCC is typically late in disease progression, the prognosis is often poor, with significant co‐morbid effects. Common risk factors include tobacco exposure, alcohol use, and human papilloma virus infection. Effects of a high fat diet (HF) on oral cancer development and severity have not been clearly defined. The purpose of this study was to compare the effects of a high saturated fat diet and a low saturated fat diet on the induction and severity of oral SCC using a mouse model of 4‐nitroquinoline 1‐oxide (4NQO) induced SCC. We hypothesized that HF would augment 4NQO‐induced SCC when compared to the LF diet.Materials and methodsMale (n=36) and female (n=36) C57Bl/6 mice (Jackson Labs), 5 weeks old, were equally distributed into a low fat (10kcal% fat; LF) or HF (60 kcal% fat) diet. Within each dietary group, mice were randomly assigned to one of 3 water treatment groups for 17 weeks: water alone (control); propylene glycol in water (1.25%; PG‐H2O); or 4NQO in PG‐H2O (50 μg/ml; 4NQO). After 17 weeks, all mice were given water alone for 6 weeks prior to euthanasia. Initial and weekly body weights were measured. Weekly assessments of the oral cavity began after 5 weeks and continued for the remaining 18 weeks. The presence of oral lesions, their size, and location were monitored and recorded weekly. Statistically significant differences in body weight, tumor size and severity were determined by ANOVA, p<0.05.ResultsAll 4‐NQO treated mice developed tumors in the oral cavity; no control mice developed oral tumors. Overall tumor burden at 23 weeks for both sexes combined was statistically higher in the HF mice compared to the LF group (10.3±1.1 vs. 7.9±0.6, p=0.055). There was no significant difference in tumor burden as a result of gender. Significant changes in weekly body weights were determined, with 4NQO mice demonstrating lower final body weights than the H2O control group and the PG‐H2O control mice on the respective diets (p<0.05). Twenty‐four hour food intake was determined at week 19, demonstrating a significant reduction in calorie intake in the 4‐NQO male mice (p<0.05).ConclusionThis mouse model of SCC, initially established to mimic the effects of smoking, demonstrated significantly greater tumor burden as a consequence of elevated fat intake. The contributory effects of HF associated metabolic and inflammatory changes in SCC development can be discerned using this model. The clinical implication of these findings may be helpful in modifying lifestyle to improve outcomes.Support or Funding InformationAcknowledgements: Andrew Nguyen was supported by the Midwestern University Biomedical Sciences Program. We also recognize the Midwestern University Deans of CDMI and the Basic Science Division for their support of this research.
Ketogenic Therapies on Cancer Cachexia in a Mouse Model of Metastatic Cancer
Authors: Koutnik Andrew P , Poff Angela M , Ward Nathan P , DeBlasi Janine , D'Agostino Dominic P 
Published: November 2023

While most cancer deaths are a result of metastatic disease, 20% directly result from cachexia. Cancer cachexia is characterized by the loss of skeletal muscle mass, with or without fat loss, that cannot be fully reversed by conventional nutritional support and results in progressive functional impairment. There are many models of cancer cachexia, but none are ideal as they do not address all aspects of this multifaceted syndrome. The VM‐M3 model is a syngeneic mouse model derived from a spontaneous brain tumor which, when implanted subcutaneously, results in rapid and systemic metastasis throughout the body, including to the liver, lungs, brain, kidneys, and spleen. Male and female VM‐M3 mice were assessed for cancer cachexia phenotype. Tumor progression, organ metastasis, skeletal muscle and adipose tissue recomposition, systemic inflammation, anorexia, albumin, blood urea nitrogen, and other clinical metabolites were evaluated. VM‐M3 male and female models both present rapid systemic metastasis (31 and 32‐day mean survival) with similar progressive increases in primary tumor weight in males and females, respectively. A progressive decrease in calf skeletal muscle weight was seen from week 2 (10% and 12% reduction) to end of life (19% and 23%), with a 40% and 26% reduction in quadriceps weight at end of life for males and females, respectively. Intraperitoneal white adipose tissue displayed progressive wasting with a 20% reduction for both male and females at week 2 and an 82% and 100% reduction for male and females, respectively. Prolonged systemic inflammation was identified via elevated spleen weight (9‐ and 10‐fold increase, respectively), elevated white blood cells counts, and elevated TNF‐α and IL‐6 at week 2 and end of life. Male and female VM‐M3 mice also presented anorexia, hypoalbumin, elevated blood urea nitrogen, and reduced total protein. Dietary interventions were developed to treat this multifaceted model of metastasis induced cancer cachexia. Ketogenic diet and synthetic ketogenic agents (R,S 1,3‐butanediol acetoacetate diester) were administered via diet to determine their ability to attenuate cachexic phenotype (altered tumor progression, skeletal muscle, adipose tissue, systemic inflammation, and additional cachexic biomarkers).Support or Funding InformationScivation Inc.
Adjuvant Statin Therapy Efficacy is Dictated by Tumor Dormancy and Statin Lipophilicity in Ex Vivo and In Vivo Models of Metastatic Breast Cancer
Authors: Beckwitt Colin H , Clark Amanda M , Warita Katsuhiko , Oltvai Zoltan N , Wells Alan 
Published: November 2023

Metastasis in breast cancer patients heralds mortality, as disseminated disease is generally chemoresistant. After tumor cells reach the ectopic tissue, they undergo an epithelial reversion to enter a period of quiescence, termed dormancy, which may last for decades before outgrowing again as mesenchymal/dedifferentiated masses. Thus, long‐term, relatively non‐toxic interventions that prevent metastatic outgrowth are needed to treat this mortal stage of tumor progression.Epidemiological analyses have suggested that statin usage, for cardiovascular indications, is correlated with a reduction in metastatic emergent (though not in incidence of primary) breast cancer. The goal of this study is to demonstrate this is due to statins suppressing breast cancer cell proliferation, a hallmark of emergent outgrowth.We have found that atorvastatin and simvastatin limit the growth of some cancer cell lines, but not others. The sensitive lines were marked by lacking surface E‐cadherin, the hallmark of the mesenchymal phenotype. When E‐cadherin is downregulated on epithelial tumor cells, the cells become growth inhibited by the statins. Furthermore, this is a direct effect, as we now have shown that hydrophilic statins are relatively ineffective compared to the membrane permeant lipophilic statins as tumor cells generally lack the transporters that enable these drugs to gain access to the cells.To determine whether the statins target the emergent metastatic tumor cells, we are using an all human microphysiological system (MPS) of the most common site for metastases, the liver. Briefly, a micro‐hepatic tissue is established by seeding primary human liver cells in a porous scaffold subject to a physiological flow. RFP‐labeled breast cancer cells are seeded into these microtissues and examined weeks later. Liver function and health are monitored by clinical chemistry assays performed on supernatant samples. We have previously shown that this system robustly reproduces tumor dormancy. Initial studies suggest that statins suppress the emergence of dormant tumor cells when challenged by stressors that lead to outgrowth. Statins were further found in vivo to influence breast cancer metastasis and metastatic growth using a mouse model of spontaneous liver metastases. As 26% of adults currently take a statin for other medical conditions, these studies may suggest the best statin to use in the context of maintaining breast cancer dormancy long‐term and delaying or avoiding the morbid emergence.Support or Funding InformationNCATS/NCI (UH3TR000496) to AW and NCI F30 (1F30CA199947), CATER Training Program (T32 EB001026), and Pitt/CMU MSTP Program (5T32GM008208) to CB
Characterization of Immunodeficient Mouse Models
Authors: McClellan Jamie , Macasocal Richard , Hare Terri , Horn Mandy 
Published: November 2023

Leakiness in SCID mouse models refers to the occasionally productive VDJ rearrangement leading to clonal expansion of these limited B and T cell clones. The rate and extent of leakiness in immunodeficient models is a factor of age as well as housing environment and the background strain of the model. In the current study, serum IgG was used to determine the extent of leakiness in various immunodeficient mouse models; survival and lymphoma incidence was also analyzed. Male and female (n=23–26) C.B‐17‐SCID, NOD SCID, SCID Beige, SHrN®, Beige Nude XID and Nude mice were housed in flexible film isolators. Blood was collected and analyzed using dried blood spot technology at 12, 16, 20 and 24 weeks of age for IgG concentration analysis using multiplex fluorescent immunoassay. The leakiness threshold was set at 5 μg/mL of IgG for analysis. At 24 weeks of age, necropsies were performed to determine incidence of lymphoma. At 24 wk of age, the SHrN® model had the smallest percentage of leaky animals followed closely by the NOD SCID. Beige Nude XID mice display an age associated increase in serum IgG levels as well as an increasing percentage of animals above the 5 μg/mL threshold from 12–24 weeks of age. Nude mice also show increases in IgG from 12 to 24 weeks of age. The NOD SCID and SHrN® mice had the lowest survival rate at 24 weeks; however, survivability in all models was greater than or equal to 79%. There is a positive correlation between the percentage of leaky animals and survival in males and females across all models. The C.B17‐SCID shows low incidence of lymphoma at 24 weeks of age whereas the NOD SCID model has the highest incidence of lymphoma but when the hairless mutation is crossed onto the NOD SCID mouse to create the SHrN®, incidence of lymphoma is decreased. This is the first study reporting IgG concentration in multiple immunodeficient models to examine leakiness as well as 24 week survival and lymphoma incidence. Additional studies measuring immune cell activity and function will be useful to further characterize immunodeficient models; however, these data will be valuable information in determining the best model for xenograft research.Support or Funding InformationThis research was privately funded by Envigo.
Molecular Diagnostic Techniques Used to Confirm an Unusual Case of BAPoma
Authors: Atkinson Aaron , Linos Konstantinos , Yan Shaofeng , Tsongalis Gregory , Lefferts Joel 
Published: November 2023

Study ObjectiveBAP1 (BRCA1‐associated protein 1) is a critical tumor suppressor that promotes genome stability and DNA damage response, in addition to other cellular processes. BAP1 maps to 3p21 and is a nuclear localized deubiquitinating hydrolase that binds BRCA1, Ino80, and Histone2A‐monoubiquitin among other proteins. Herein, we employ advanced molecular diagnostic techniques to confirm the suspected diagnosis of a BAP1‐deficient Spitzoid Tumor (BAPoma) following confounding stereotypical histology and immunohistochemical tests. Proper assessment of Bap1‐deficient tumors is important as the likelihood of tumor progression if left untreated is currently unknown, however in other cancers such as uveal melanoma BAP1‐deficient tumors are treated as higher risk.Clinical Presentation and MethodsA 45 year old female patient presented an irregular translucent papule on her shoulder that when biopsied had hallmarks of a BAPoma including: biphasic melanocytic proliferation, and immunostaining showing BRAF V600E; p16 expression was retained. However, nuclear BAP1 immunohistochemistry was maintained. To confirm the diagnosis of BAPoma, we performed Chromosomal Microarray (CMA) analysis using Oncoscan FFPE Assay (Affymetrix); Sanger sequencing of BAP1 commonly mutated regions; and SNaPShot primer‐extension based genotyping.ResultsDNA was extracted from the tissue biopsy and used in CMA testing that revealed a heterozygous loss of 3p. To reconcile the morphologic features with both the nuclear BAP1 staining and loss of 3p, Sanger sequencing of exons 4–7 and 17 PCR products was performed. Sequencing revealed a suspected c.505C>T p.H196Y mutation that was also confirmed by primer‐specific mini‐sequencing. This BAP1 variant is likely nonfunctional and pathogenic as, in addition to H169 being among 4 conserved catalytic residues, known pathogenic H169 substitutions, including H169Y, have been found previously in uveal melanoma, mesothelioma, and bladder cancer.ConclusionsThe presence of a nonfunctional allele within the nucleus combined with loss of BAP1 heterozygosity explains the clear and characteristic BAPoma morphology by histology. While such cases are rare, this case underlines the importance of molecular diagnostics when protein expression studies do not correlate with morphology. Future molecular testing could be refined to target known missense mutations in BAP1 that do not affect nuclear localization ‐ especially those in catalytic residues such as H169.