tbuckley/cabot-search huggingface dataset:
Cilastatin protects against imipenem-induced nephrotoxicity via inhibition of renal organic anion transporters (OATs)
Authors: Xiaokui Huo, Qiang Meng, Changyuan Wang, Yanna Zhu, Zhihao Liu, Xiaodong Ma, Xiaochi Ma, Jinyong Peng, Huijun Sun, Kexin Liu
Journal: Acta Pharmaceutica Sinica B (2019)

Imipenem is a carbapenem antibiotic. However, Imipenem could not be marketed owing to its instability and nephrotoxicity until cilastatin, an inhibitor of renal dehydropeptidase-I (DHP-I), was developed. In present study, the potential roles of renal organic anion transporters (OATs) in alleviating the nephrotoxicity of imipenem by cilastatin were investigated in vitro and in rabbits. Our results indicated that imipenem and cilastatin were substrates of hOAT1 and hOAT3. Cilastatin inhibited hOAT1/3-mediated transport of imipenem with IC50 values comparable to the clinical concentration, suggesting the potential to cause a clinical drug-drug interaction (DDI). Moreover, imipenem exhibited hOAT1/3-dependent cytotoxicity, which was alleviated by cilastatin and probenecid. Furthermore, cilastatin and probenecid ameliorated imipenem-induced rabbit acute kidney injury, and reduced the renal secretion of imipenem. Cilastatin and probenecid inhibited intracellular accumulation of imipenem and sequentially decreased the nephrocyte toxicity in rabbit primary proximal tubule cells. Renal OATs, besides DHP-I, was also the target of interaction between imipenem and cilastatin, and contributed to the nephrotoxicity of imipenem. This therefore gives in part the explanation about the mechanism by which cilastatin protected against imipenem-induced nephrotoxicity. Thus, OATs can potentially be used as a therapeutic target to avoid the renal adverse reaction of imipenem in clinic.
P110/O18 Epigenetic changes by inhibition of DOT1L affect wnt signaling, proliferation and cell cycle in dermal fibroblasts, with no overall effect on collagen deposition in models of fibrosis
Authors: N Berghen, J Cremer, E De Langhe, R Lories
Journal: Annals of the Rheumatic Diseases (2019)

<h3>Career situation of first and presenting author</h3> Student for a master or a PhD. <h3>Introduction</h3> The role of epigenetic factors in the pathophysiology of fibrosis, a hallmark of Systemic Sclerosis, is increasingly explored. DOT1L, the unique H3K79-methyltransferase, methylates histone 3 at the Lysine residue at position 79, thereby regulating gene expression programs. In cartilage and bone, DOT1L has cell-type specific effects on Wnt signaling, a pathway suggested to play an important role in fibrosis. <h3>Objectives</h3> To study the role of DOT1L in fibrosis. <h3>Methods</h3> Primary human dermal fibroblasts were treated with DOT1L-inhibitor EPZ-5676 or vehicle and stimulated with TGF-β. Expression of smooth muscle alpha 2 actin (ACTA2) and Wnt target genes was measured by RT-qPCR. Western Blot was done for dimethylated H3K79 and β-catenin. Picrosirius Red staining measured collagen deposition. 5-Bromo-2'-deoxy-uridine (BrdU) labeling for proliferation and flow cytometry with Propidium Iodide for cell cycle analysis was done. Col1a2;Cre-ERT<sup>2</sup>;DOT1l<sup>fl/fl</sup>mice, injected with tamoxifen to induce a fibroblast-specific DOT1L knockout, were injected subcutaneously with bleomycin or vehicle. Injected skin was analyzed by OH-prolin assay and by histology. <h3>Results</h3> The DOT1L-inhibitor EPZ-5676 reduced H3K79 dimethylation in all samples. In breast dermal fibroblasts, the induction of ACTA2 with TGF-β was reduced with DOT1L inhibition, while in abdominal dermal fibroblasts this induction was more pronounced. After 48 hours of TGF-β, collagen deposition was higher in DOT1L-inhibited fibroblasts. After 72 hours of TGF-β however, this deposition was comparable with controls. DOT1L inhibition induced canonical Wnt signaling in fibroblasts, with a small increase in active β-catenin and expression of Lymphoid enhancer-binding factor 1 (LEF1). With DOT1L inhibition, more proliferation of fibroblasts, but also proportionally more cells in the G1/G0 phase and less cells in S and M/G2 phase were seen. <i>In vivo</i>, subcutaneous bleomycin increased murine dermal thickness and skin collagen content. No difference was observed between wild type and mice with a fibroblast-specific deletion of DOT1L. <h3>Conclusions</h3> In an <i>in vitro</i> model of fibrosis, ACTA2 induction in DOT1L-inhibited human dermal fibroblasts was dependent on the fibroblast origin. DOT1L inhibition resulted in an earlier deposition of collagen, without differences in deposition at the end points. Inhibition of DOT1L induced canonical Wnt signaling and proliferation but also led to a higher proportion of cells in the G0/G1 phase. In an <i>in vivo</i> murine model of skin fibrosis, no difference in bleomycin-induced skin thickness and collagen content was found when the DOT1L gene was deleted in fibroblasts. <h3>Disclosure of Interest</h3> None declared.
Abstract 2540: Identification of factors governing sensitivity of NSCLC cell lines to inhibition of Poly (ADP-Ribose) Polymerase (PARP)
Authors: Naveen Kommajosyula, Lisa Moreau, Alan D. D'Andrea, Geoffrey I. Shapiro
Journal: Cancer Research (2012)

Abstract Recently, Poly (ADP Ribose) Polymerase-1 (PARP-1) has been shown to be an important target in subsets of breast and ovarian cancers. PARP inhibitors are emerging as important new anticancer agents, especially in cancers defective in homologous recombination (HR) repair, such as those lacking BRCA1 or BRCA2. To date, the potential role of PARP inhibition in NSCLC has not been extensively studied. BRCA1 or BRCA2 promoter hypermethylation and ATM mutation have been described in primary NSCLCs, alterations that may sensitize cells to PARP inhibition. We have examined a panel of 26 genotypically-defined NSCLC cell lines for sensitivity to the PARP-1 inhibitor AG014699 using colony formation assays. We have identified 5cell lines that are exquisitely sensitive (IC50 ≤ 50nM), 9 cell lines that are moderately sensitive (IC50 ≤ 60-500nM) and 12 cell lines that are resistant (IC50 &amp;gt; 5 µM). To determine whether sensitive cell lines have defective HR compared to resistant cell lines, we examined the formation of Rad51 foci following exposure to PARP inhibition, as well as to gamma-irradiation and cisplatin. Both sensitive and resistant cell lines demonstrated efficient formation of Rad51 foci as well as gamma-H2AX foci in response to DNA damaging treatments, suggesting correct initiation of HR. In resistant cell lines, the repair process following PARP inhibition is completed within 24 hours, with resolution of Rad51 and gamma H2AX foci. No alterations in cell cycle distribution were detected in these cell lines, suggesting that the efficiency of repair allows proliferation to continue largely unabated. However, in sensitive cell lines, these foci persist greater than 30 hours following DNA damaging treatments. These results suggest that although sensitive lines are able to initiate HR repair, they are unable to complete the process, so that Rad51 foci are not resolved. The persistence of DNA damage in these cell lines is confirmed by persistent gamma H2AX focus formation as well as S/G2 arrest, the latter representing intact checkpoint control. We are currently investigating whether PARP inhibitor-sensitive cell lines have a deficiency in one of the helicases or resolvases required for resolution of the complex Holliday junctions formed during HR repair. Our results may define a novel subset of NSCLCs with a late-step defect in HR that can be approached with PARP inhibition as a novel treatment strategy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2540. doi:1538-7445.AM2012-2540
Intranasal Instilation of a Very Low Dose of ROFA Compromises Lung Mechanics and Histology for 4 Days
Authors: Walter Zin, Lilian Nagato, Sheila Fagundes, Andrea Calheiros, Patrícia Bozza, Olaf Malm, Paulo H. N. Saldiva, Debora Faffe
Journal: European Respiratory Journal (2007)
Mechanical stress-induced mast cell degranulation activates TGF-β1 signalling pathway in pulmonary fibrosis
Authors: Chiko Shimbori, Chandak Upagupta, Pierre-Simon Bellaye, Ehab A Ayaub, Seidai Sato, Toyoshi Yanagihara, Quan Zhou, Alexander Ognjanovic, Kjetil Ask, Jack Gauldie, Paul Forsythe, Martin R J Kolb
Journal: Thorax (2019)

Background The role of mast cells accumulating in idiopathic pulmonary fibrosis (IPF) lungs is unknown. Objectives We investigated the effect of fibrotic extracellular matrix (ECM) on mast cells in experimental and human pulmonary fibrosis. Results In IPF lungs, mast cell numbers were increased and correlated with disease severity (control vs 60%&lt;FVC&lt;90%, mean difference=-222.7, 95% CI −386.3 to −59.2, p=0.004; control vs FVC&lt;60%, mean difference=−301.7, 95% CI of difference −474.1 to −129.34, p=0.0001; FVC&gt;90% vs 60%&lt;FVC&lt;90%, mean difference=−189.6, 95% CI of difference −353.1 to −26.03, p=0.017; FVC&gt;90% vs FVC&lt;60%, mean difference=−268.6, 95% CI of difference −441.0 to −96.17, p=0.0007). Plasma tryptase levels were increased in IPF and negatively correlated with FVC (control vs FVC&lt;60%, mean difference=−17.12, 95% CI of difference −30.02 to −4.22, p=0.006: correlation curves R=−0.045, p=0.025). In a transforming growth factor (TGF)-β1-induced pulmonary fibrosis model, chymase-positive and tryptase-positive mast cells accumulated in fibrotic lung. Lung tissue was decellularised and reseeded with bone marrow or peritoneum-derived mast cells; cells on fibrotic ECM released more TGF-β1 compared with normal ECM (active TGF-β1: bone marrow-derived mast cell (BMMC)-DL vs BMMC-TGF-β1 p=0.0005, peritoneal mast cell (PMC)-DL vs PMC-TGF-β1 p=0.0003, total TGF-β1: BMMC-DL vs BMMC-TGF-β1 p=0.013, PMC-DL vs PMC-TGF-β1 p=0.001). Mechanical stretch of lungs caused mast cell degranulation; mast cell stabilisers inhibited degranulation (histamine: cont vs doxantrazole p=0.004, β-hexosaminidase: cont vs doxantrazole, mean difference=1.007, 95% CI of difference 0.2700 to 1.744, p=0.007) and TGF-β1 activation (pSmad2/Smad2: cont vs dox p=0.006). Cromoglycate attenuated pulmonary fibrosis in rats (collagen: phosphate-buffered saline (PBS) vs cromoglycate p=0.036, fibrotic area: PBS vs cromoglycate p=0.031). Conclusion This study suggests that mast cells may contribute to the progression of pulmonary fibrosis.
Phase I Immunotherapy Trial with Two Chimeric HER-2 B-Cell Peptide Vaccines Emulsified in Montanide ISA 720VG and Nor-MDP Adjuvant in Patients with Advanced Solid Tumors
Authors: Tanios Bekaii-Saab, Robert Wesolowski, Daniel H. Ahn, Christina Wu, Amir Mortazavi, Maryam Lustberg, Bhuvaneswari Ramaswamy, Jeffrey Fowler, Lai Wei, Jay Overholser, Pravin T.P. Kaumaya
Journal: Clinical Cancer Research (2019)

This first-in-human phase I study (NCT01417546) evaluated the safety profile, optimal immunologic/biological dose (OID/OBD), and immunogenicity of the combination of two peptide B-cell epitope vaccines engineered to represent the trastuzumab- and pertuzumab-binding sites. Although trastuzumab and pertuzumab have been approved for clinical use, patients often develop resistance to these therapies. We have advanced a new paradigm in immunotherapy that focuses on humoral responses based on conformational B-cell epitope vaccines.The vaccine is comprised of two chimeric HER-2 B-cell peptide vaccines incorporating a "promiscuous T-cell epitope." Patients were immunized with the vaccine constructs emulsified with nor-muramyl-dipeptide adjuvant in a water-in-oil Montanide ISA 720VG vehicle. Eligible patients with metastatic and/or recurrent solid tumors received three inoculations every 3 weeks.Forty-nine patients with a median of 4 prior lines of chemotherapy received at least 1 vaccination. Twenty-eight patients completed the 3 vaccination regimens. Six patients received 1 six-month boost after the regimen, and one patient received 7 six-month boosts. No serious adverse reactions or dose-limiting toxicities were observed. The vaccine was well tolerated with dose level 2 as the recommended phase II dose. The most common related toxicity in all patients was injection-site reactions (24%). Two patients had a partial response, 14 had stable disease, and 19 had progressive disease.The study vaccine is safe, exhibits antitumor activity, and shows preliminary indication that peptide vaccination may avoid therapeutic resistance and offer a promising alternative to monoclonal antibody therapies.
On-target Resistance to the Mutant-Selective EGFR Inhibitor Osimertinib Can Develop in an Allele-Specific Manner Dependent on the Original EGFR-Activating Mutation
Authors: Benjamin P. Brown, Yun-Kai Zhang, David Westover, Yingjun Yan, Huan Qiao, Vincent Huang, Zhenfang Du, Jarrod A. Smith, Jeffrey S. Ross, Vincent A. Miller, Siraj Ali, Lyudmila Bazhenova, Alexa B. Schrock, Jens Meiler, Christine M. Lovly
Journal: Clinical Cancer Research (2019)

The third-generation EGFR inhibitor, osimertinib, is the first mutant-selective inhibitor that has received regulatory approval for the treatment of patients with EGFR-mutant lung cancer. Despite the development of highly selective third-generation inhibitors, acquired resistance remains a significant clinical challenge. Recently, we and others have identified a novel osimertinib resistance mutation, G724S, which was not predicted in in vitro screens. Here, we investigate how G724S confers resistance to osimertinib.Experimental Design: We combine structure-based predictive modeling of G724S in combination with the 2 most common EGFR-activating mutations, exon 19 deletion (Ex19Del) and L858R, with in vitro drug-response models and patient genomic profiling.Our simulations suggest that the G724S mutation selectively reduces osimertinib-binding affinity in the context of Ex19Del. Consistent with our simulations, cell lines transduced with Ex19Del/G724S demonstrate resistance to osimertinib, whereas cells transduced with L858R/G724S are sensitive to osimertinib. Subsequent clinical genomic profiling data further suggest G724S occurs with Ex19Del but not L858R. Furthermore, we demonstrate that Ex19Del/G724S retains sensitivity to afatinib, but not to erlotinib, suggesting a possible therapy for patients at the time of disease relapse.Altogether, these data suggest that G724S is an allele-specific resistance mutation emerging in the context of Ex19Del but not L858R. Our results fundamentally reframe the problem of targeted therapy resistance from one focused on the "drug-resistance mutation" pair to one focused on the "activating mutation-drug-resistance mutation" trio. This has broad implications across clinical oncology.
Comment on Warren et al. Diabetes and Trajectories of Estimated Glomerular Filtration Rate: A Prospective Cohort Analysis of the Atherosclerosis Risk in Communities Study. Diabetes Care 2018;41:1646–1653
Authors: Ying-Xiu Kang, Xi-Ling Lin, Yue Ding, Xiao-Wen Pan, Shu-Xia He, Peng-Fei Shan
Journal: Diabetes Care (2019)

In the study by Warren et al. (1), the authors reported on long-term kidney disease trajectories in persons with and without diabetes from four U.S. communities. They concluded that kidney function in those with diabetes declined almost twice as rapidly as it did in those without diabetes. This was a large-scale, well-conducted, prospective clinical cohort analysis including 15,517 participants at four visits, lasting over 26 years. The study provided precious clinical data on changes in renal function in patients with diabetes over many years. However, we would like to discuss an issue related to the deterioration rate of kidney function in subjects with different estimated glomerular filtration rate (eGFR) values.

In the study by Warren et al. (1), the mean annual decline in eGFR was …
Protein Abnormalities in Neuromuscular Diseases—Part 1
Authors: W. King Engel
Journal: JAMA (1966)

<h3>Dr. W. King Engel (Moderator):</h3> We will begin the discussion of protein abnormalities in various neuromuscular diseases by first explaining our use of the term "neuromuscular diseases." Figure 1 is a diagram of what is called the "motor unit." The anterior-horn motor neuron (or lower motor neuron) lies in the anterior horn of the spinal cord, and the peripheral extension of that neuron goes out through the anterior root, through the peripheral nerve, to the terminal branches of that nerve cell. At the end of the terminal branches are the neuromuscular junctions, the connections between the nerve cell and the numerous muscle fibers, generally several hundred, that one neuron innervates. A motor neuron and the muscle fibers innervated by it comprise a "motor unit." Diseases that affect any part of the motor unit—that is, the anterior horn cell, peripheral nerve, neuromuscular junction, or muscle—are termed "neuromuscular diseases." In various conditions,
Ferroptotic cell death and TLR4/Trif signaling initiate neutrophil recruitment after heart transplantation
Authors: Wenjun Li, Guoshuai Feng, Jason M. Gauthier, Inessa Lokshina, Ryuji Higashikubo, Sarah Evans, Xinping Liu, Adil Hassan, Satona Tanaka, Markus Cicka, Hsi-Min Hsiao, Daniel Ruiz-Perez, Andrea Bredemeyer, Richard W. Gross, Douglas L. Mann, Yulia Y. Tyurina, Andrew E. Gelman, Valerian E. Kagan, Andreas Linkermann, Kory J. Lavine, Daniel Kreisel
Journal: Journal of Clinical Investigation (2019)

Nonapoptotic forms of cell death can trigger sterile inflammation through the release of damage-associated molecular patterns, which are recognized by innate immune receptors. However, despite years of investigation, the mechanisms that initiate inflammatory responses after heart transplantation remain elusive. Here, we demonstrate that ferrostatin-1 (Fer-1), a specific inhibitor of ferroptosis, decreases the levels of pro-ferroptotic hydroperoxy-arachidonoyl-phosphatidylethanolamine, reduces cardiomyocyte cell death, and blocks neutrophil recruitment following heart transplantation. Inhibition of necroptosis had no effect on neutrophil trafficking in cardiac grafts. We extend these observations to a model of coronary artery ligation–induced myocardial ischemia reperfusion injury (IRI), in which inhibition of ferroptosis resulted in reduced infarct size, improved left ventricular (LV) systolic function, and reduced LV remodeling. Using intravital imaging of cardiac transplants, we show that ferroptosis orchestrates neutrophil recruitment to injured myocardium by promoting adhesion of neutrophils to coronary vascular endothelial cells through a TLR4/Trif/type I IFN signaling pathway. Thus, we have discovered that inflammatory responses after cardiac transplantation are initiated through ferroptotic cell death and TLR4/Trif-dependent signaling in graft endothelial cells. These findings provide a platform for the development of therapeutic strategies for heart transplant recipients and patients who are vulnerable to IRI following restoration of coronary blood flow.
Abstract 11232: Arterial Stiffness is an Independent Predictor of Cardiac Mortality in Asymptomatic Patients With Intermediate Atherosclerotic Risk Factors
Authors: Teerapat Yingchoncharoen, Thosaphol Limpijankit, Sutipong Jongjirasiri, Jiraporn Laothamatas, Piyamitr Sritara
Journal: Circulation (2013)

Background: To identify predictors of cardiac death in patients with intermediate atherosclerotic risk patients, we studied the prognostic value of arterial stiffness as measured by Cardio Ankle Vascular Index (CAVI) in these patients. Methods: We prospectively followed 1,382 asymptomatic patients with intermediate risk factors as assessed by RAMA-EGAT score showing predicted 10-years event rate of 10-20%. RAMA-EGAT Score is a Framingham risk score equivalent but more well-validated in Thai population. These patients were referred for Coronary Computed Tomography Angiography (CTA) for cardiac risk assessment. Patients were followed over 6.9+0.7 years for cardiac death (54 patients). A multivariable Cox regression was performed to identify associations with events. Results: Event free survival was 96+0.5% at 6 years. Cardiac mortality was predicted by CAVI (Hazard ratio 1.23 [95%CI 1.01-1.54], p = 0.046), independent of traditional risk score validated in our population (RAMA-EGAT Score) (Hazard ratio 1.01...
P069 Constitutive overexpression of interleukin 38 has a negative impact on human NHK keratinocyte fitness
Authors: L Mermoud, D Talabot-Ayer, N Brembilla, J Julia Borowczyk-Michalowska, G Palmer
Journal: Annals of the Rheumatic Diseases (2019)

<h3>Career situation of first and presenting author</h3> Student for a master or a PhD. <h3>Introduction</h3> Interleukin (IL)-38 is a member of the IL-1 cytokine family. Based on its sequence homology with the IL-1 and IL-36 receptor antagonists, IL-38 was proposed to act as an anti-inflammatory molecule. However, few data are available to date about its biological role and mechanism of action. Highest constitutive IL-38 expression is detected in the skin, where the cytokine is produced mainly by differentiated keratinocytes. <h3>Objectives</h3> The aim of this study is to investigate the function of IL-38 in human keratinocytes, using the immortalized normal human keratinocyte (NHK) cell line stably transfected with vectors allowing for constitutive or inducible IL-38 overexpression. <h3>Methods</h3> NHK cell proliferation and mortality were assessed respectively by cell counting and by measuring LDH activity in the supernatant of high density cell culture. Differentiation of NHK cells was initiated by switching to a culture medium containing high Ca<sup>++</sup> and serum. mRNA levels of differentiating NHK cells were determined by RTqPCR. <h3>Results</h3> Constitutive IL-38 overexpression reduced the proliferation of NHK cells and lead to increased mortality. Differentiating NHK cells constitutively overexpressing IL-38 had increased mRNA levels for the early differentiation marker keratin 10 as compared to empty vector transfected control cells. On the contrary, expression of the late differentiation marker involucrin was reduced in NHK cells constitutively overexpressing IL-38. To further explore potential effects of IL-38 on keratinocyte differentiation, while circumventing the impact of its constitutive overexpression on cell fitness, we set up a doxycyclin-inducible Tet-On system to overexpress IL-38 in NHK cells. <h3>Conclusions</h3> We are currently optimizing 2D and 3D culture conditions for <i>in vitro</i> differentiation of NHK cells to mimic more closely the situation in human skin. We will then study the impact of IL-38 on NHK cell differentiation using cells transfected with the inducible IL-38 expression system. In conclusion, our results suggest that constitutive overexpression of IL-38 decreases proliferation and viability of NHK cells. We will thus use an inducible expression system to further investigate the effects of IL-38 on NHK cells. <h3>Disclosure of Interest</h3> None declared.
TP53 Mutations Negatively Impact Survival of Acute Myeloid Leukemia Patients Treated with Standard Doses of Azacitidine
Authors: Pierre Bories, Naïs Prade, Stéphanie Lagarde, Bastien Cabarrou, Julien Plenecassagnes, Isabelle Luquet, Veronique De Mas, Thomas Filleron, Manon Cassou, Audrey Sarry, Luc Matthieu Fornecker, Célestine Simand, Sarah Bertoli, Hinrich Gronemeyer, Christian Recher, Eric Delabesse
Journal: Blood (2018)