30 results on '"Amrita Cheema"'
Search Results
2. Membrane Stress Caused by Unprocessed Outer Membrane Lipoprotein Intermediate Pro-Lpp Affects DnaA and Fis-Dependent Growth
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Digvijay Patil, Dan Xun, Markus Schueritz, Shivani Bansal, Amrita Cheema, Elliott Crooke, and Rahul Saxena
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Escherichia coli ,DNA replication ,acidic phospholipids ,lipoprotein biogenesis ,DnaA ,Fis ,Microbiology ,QR1-502 - Abstract
In Escherichia coli, repression of phosphatidylglycerol synthase A gene (pgsA) lowers the levels of membrane acidic phospholipids, particularly phosphatidylglycerol (PG), causing growth-arrested phenotype. The interrupted synthesis of PG is known to be associated with concomitant reduction of chromosomal content and cell mass, in addition to accumulation of unprocessed outer membrane lipoprotein intermediate, pro-Lpp, at the inner membrane. However, whether a linkage exists between the two altered-membrane outcomes remains unknown. Previously, it has been shown that pgsA+ cells overexpressing mutant Lpp(C21G) protein have growth defects similar to those caused by the unprocessed pro-Lpp intermediate in cells lacking PG. Here, we found that the ectopic expression of DnaA(L366K) or deletion of fis (encoding Factor for Inversion Stimulation) permits growth of cells that otherwise would be arrested for growth due to accumulated Lpp(C21G). The DnaA(L366K)-mediated restoration of growth occurs by reduced expression of Lpp(C21G) via a σE-dependent small-regulatory RNA (sRNA), MicL-S. In contrast, restoration of growth via fis deletion is only partially dependent on the MicL-S pathway; deletion of fis also rescues Lpp(C21G) growth arrest in cells lacking physiological levels of PG and cardiolipin (CL), independently of MicL-S. Our results suggest a close link between the physiological state of the bacterial cell membrane and DnaA- and Fis-dependent growth.
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- 2021
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3. Publisher Correction: Hypoxia-activated neuropeptide Y/Y5 receptor/RhoA pathway triggers chromosomal instability and bone metastasis in Ewing sarcoma
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Congyi Lu, Akanksha Mahajan, Sung-Hyeok Hong, Susana Galli, Shiya Zhu, Jason U. Tilan, Nouran Abualsaud, Mina Adnani, Stacey Chung, Nada Elmansy, Jasmine Rodgers, Olga Rodriguez, Christopher Albanese, Hongkun Wang, Maureen Regan, Valerie Zgonc, Jan Blancato, Ewa Krawczyk, G. Ian Gallicano, Michael Girgis, Amrita Cheema, Ewa Iżycka-Świeszewska, Luciane R. Cavalli, Svetlana D. Pack, and Joanna Kitlinska
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Science - Published
- 2022
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4. Exercise modifies glutamate and other metabolic biomarkers in cerebrospinal fluid from Gulf War Illness and Myalgic encephalomyelitis / Chronic Fatigue Syndrome.
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James N Baraniuk, Grant Kern, Vaishnavi Narayan, and Amrita Cheema
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Medicine ,Science - Abstract
Myalgic encephalomyelitis / Chronic Fatigue Syndrome (ME/CFS) and Gulf War Illness (GWI) share many symptoms of fatigue, pain, and cognitive dysfunction that are not relieved by rest. Patterns of serum metabolites in ME/CFS and GWI are different from control groups and suggest potential dysfunction of energy and lipid metabolism. The metabolomics of cerebrospinal fluid was contrasted between ME/CFS, GWI and sedentary controls in 2 sets of subjects who had lumbar punctures after either (a) rest or (b) submaximal exercise stress tests. Postexercise GWI and control subjects were subdivided according to acquired transient postexertional postural tachycardia. Banked cerebrospinal fluid specimens were assayed using Biocrates AbsoluteIDQ® p180 kits for quantitative targeted metabolomics studies of amino acids, amines, acylcarnitines, sphingolipids, lysophospholipids, alkyl and ether phosphocholines. Glutamate was significantly higher in the subgroup of postexercise GWI subjects who did not develop postural tachycardia after exercise compared to nonexercise and other postexercise groups. The only difference between nonexercise groups was higher lysoPC a C28:0 in GWI than ME/CFS suggesting this biochemical or phospholipase activities may have potential as a biomarker to distinguish between the 2 diseases. Exercise effects were suggested by elevation of short chain acylcarnitine C5-OH (C3-DC-M) in postexercise controls compared to nonexercise ME/CFS. Limitations include small subgroup sample sizes and absence of postexercise ME/CFS specimens. Mechanisms of glutamate neuroexcitotoxicity may contribute to neuropathology and "neuroinflammation" in the GWI subset who did not develop postural tachycardia after exercise. Dysfunctional lipid metabolism may distinguish the predominantly female ME/CFS group from predominantly male GWI subjects.
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- 2021
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5. Safety and Pharmacokinetic Assessment of Oral Proglumide in Those with Hepatic Impairment
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Christine C. Hsu, Sunil Bansal, Hong Cao, Coleman I. Smith, Aiwu Ruth He, Martha D. Gay, Yaoxiang Li, Amrita Cheema, and Jill P. Smith
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cholecystokinin receptor ,cirrhosis ,hepatocellular carcinoma ,pharmacokinetics ,Pharmacy and materia medica ,RS1-441 - Abstract
Proglumide is an orally administered cholecystokinin receptor antagonist that was found to improve nonalcoholic steatohepatitis, reverse liver fibrosis, and decrease incidence of hepatocellular carcinoma (HCC) in animal models. The current investigation aimed to test the pharmacokinetics and safety of proglumide in subjects with hepatic impairment compared with healthy controls. In this translational study, subjects with confirmed cirrhosis, Child-Pugh stage A or B, or healthy controls were recruited for a single-dosing study. Baseline urine and blood samples were obtained before administration of proglumide and also collected after ingestion up to 24 h. Drug concentrations measured by mass spectroscopy revealed peak plasma concentrations (Cmax) of 7847, 9721, and 10,635 ng/mL at about 1 h (Tmax) for healthy controls, subjects with Child-Pugh A, and B cirrhosis, respectively. The serum elimination half time was 3 h. Maximum urine drug concentration (Cmax = ~411 µg/mL) was observed at 3 h, and urinary drug concentration declined at 5 h. There were no adverse events reported, and follow-up liver panels in cirrhosis subjects were unchanged or improved. This investigation demonstrated that proglumide is safe and has similar pharmacokinetic properties in subjects with cirrhosis as in healthy controls; therefore, it will be safe to test the efficacy of proglumide as a therapeutic agent in those subjects with cirrhosis or HCC.
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- 2022
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6. Proteomic profiles for Alzheimer's disease and mild cognitive impairment among adults with Down syndrome spanning serum and plasma: An Alzheimer's Biomarker Consortium–Down Syndrome (ABC–DS) study
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Melissa E. Petersen, Fan Zhang, Nicole Schupf, Sharon J. Krinsky‐McHale, James Hall, Mark Mapstone, Amrita Cheema, Wayne Silverman, Ira Lott, Michael S. Rafii, Benjamin Handen, William Klunk, Elizabeth Head, Brad Christian, Tatiana Foroud, Florence Lai, H. Diana Rosas, Shahid Zaman, Beau M. Ances, Mei‐Cheng Wang, Benjamin Tycko, Joseph H. Lee, Sid O'Bryant, and the Alzheimer's Biomarker Consortium – Down Syndrome (ABC‐DS)
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Neurology. Diseases of the nervous system ,RC346-429 ,Geriatrics ,RC952-954.6 - Abstract
Abstract Introduction Previously generated serum and plasma proteomic profiles were examined among adults with Down syndrome (DS) to determine whether these profiles could discriminate those with mild cognitive impairment (MCI‐DS) and Alzheimer's disease (DS‐AD) from those cognitively stable (CS). Methods Data were analyzed on n = 305 (n = 225 CS; n = 44 MCI‐DS; n = 36 DS‐AD) enrolled in the Alzheimer's Biomarker Consortium–Down Syndrome (ABC–DS). Results Distinguishing MCI‐DS from CS, the serum profile produced an area under the curve (AUC) = 0.95 (sensitivity [SN] = 0.91; specificity [SP] = 0.99) and an AUC = 0.98 (SN = 0.96; SP = 0.97) for plasma when using an optimized cut‐off score. Distinguishing DS‐AD from CS, the serum profile produced an AUC = 0.93 (SN = 0.81; SP = 0.99) and an AUC = 0.95 (SN = 0.86; SP = 1.0) for plasma when using an optimized cut‐off score. AUC remained unchanged to slightly improved when age and sex were included. Eotaxin3, interleukin (IL)‐10, C‐reactive protein, IL‐18, serum amyloid A , and FABP3 correlated fractions at r2 > = 0.90. Discussion Proteomic profiles showed excellent detection accuracy for MCI‐DS and DS‐AD.
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- 2020
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7. Treatment With Tetrahydrobiopterin Improves White Matter Maturation in a Mouse Model for Prenatal Hypoxia in Congenital Heart Disease
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Jennifer Romanowicz, Camille Leonetti, Zaenab Dhari, Ludmila Korotcova, Shruti D. Ramachandra, Nemanja Saric, Paul D. Morton, Shivani Bansal, Amrita Cheema, Vittorio Gallo, Richard A. Jonas, and Nobuyuki Ishibashi
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congenital heart disease ,hypoxia ,neuroprotection ,tetrahydrobiopterin ,Diseases of the circulatory (Cardiovascular) system ,RC666-701 - Abstract
Background Reduced oxygen delivery in congenital heart disease causes delayed brain maturation and white matter abnormalities in utero. No treatment currently exists. Tetrahydrobiopterin (BH4) is a cofactor for neuronal nitric oxide synthase. BH4 availability is reduced upon NOS activation, such as during hypoxic conditions, and leads to toxin production. We hypothesize that BH4 levels are depleted in the hypoxic brain and that BH4 replacement therapy mitigates the toxic effects of hypoxia on white matter. Methods and Results Transgenic mice were used to visualize oligodendrocytes. Hypoxia was introduced during a period of white matter development equivalent to the human third trimester. BH4 was administered during hypoxia. BH4 levels were depleted in the hypoxic brain by direct quantification (n=7–12). The proliferation (n=3–6), apoptosis (n=3–6), and developmental stage (n=5–8) of oligodendrocytes were determined immunohistologically. Total oligodendrocytes increased after hypoxia, consistent with hypoxia‐induced proliferation seen previously; however, mature oligodendrocytes were less prevalent in hypoxia, and there was accumulation of immature oligodendrocytes. BH4 treatment improved the mature oligodendrocyte number such that it did not differ from normoxia, and accumulation of immature oligodendrocytes was not observed. These results persisted beyond the initial period of hypoxia (n=3–4). Apoptosis increased with hypoxia but decreased with BH4 treatment to normoxic levels. White matter myelin levels decreased following hypoxia by western blot. BH4 treatment normalized myelination (n=6–10). Hypoxia worsened sensory‐motor coordination on balance beam tasks, and BH4 therapy normalized performance (n=5–9). Conclusions Suboptimal BH4 levels influence hypoxic white matter abnormalities. Repurposing BH4 for use during fetal brain development may limit white matter dysmaturation in congenital heart disease.
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- 2019
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8. 68477 Pancreatic cancer cell extracellular vesicles drive the unfolded protein response in recipient normal pancreatic cells.
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Charles Hinzman, Shivani Bansal, Yaoxiang Li, Jose Trevino, Partha Banerjee, and Amrita Cheema
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Medicine - Abstract
ABSTRACT IMPACT: This study advances our understanding of potentially key drivers in the early formation of pancreatic cancer, a disease with few treatment options and poor patient outcomes. OBJECTIVES/GOALS: Patients diagnosed with pancreatic ductal adenocarcinoma (PDAC) have a 5-year survival rate of ˜9%. A key driver of poor patient outcomes is late-stage diagnosis. A better understanding of PDAC onset is needed. This study was developed to understand how extracellular vesicles may be involved in the early formation of PDAC. METHODS/STUDY POPULATION: Extracellular vesicles (EVs) were isolated from several human PDAC and normal pancreatic cell lines, using ultracentrifugation with filtration or size exclusion chromatography. We next treated normal pancreatic cell lines with cancer cell EVs (cEVs). Next generation sequencing was used to measure global gene expression changes after treatment. Validations were performed using qPCR and luciferase activity assays. Multi-omics characterization of EVs was accomplished using mass spectrometry based proteomics, metabolomics and lipidomics analysis. RESULTS/ANTICIPATED RESULTS: We found that normal cells upregulated a variety of stress response pathways in response to cEVs. Lipid synthesis was also severely downregulated in these cells. We further validated activation of the unfolded protein response (UPR) in normal cells treated with cEVs. Multi-omics characterization of cEVs identified several enriched proteins, lipids and metabolites which may play a role in the activation of the UPR. DISCUSSION/SIGNIFICANCE OF FINDINGS: Our results indicate that cEVs induce stress, and in particular the UPR, in normal pancreatic cells. Long-term UPR can impact a variety of cancer hallmarks. The UPR can mediate progression of pancreatic intraepithelial neoplasia (PanIN) to PDAC. Our results highlight a potential role for cEVs to alter the function of normal cells, aiding disease onset.
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- 2021
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9. Multi-Omic Analysis Reveals Different Effects of Sulforaphane on the Microbiome and Metabolome in Old Compared to Young Mice
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Se-Ran Jun, Amrita Cheema, Chhanda Bose, Marjan Boerma, Philip T. Palade, Eugenia Carvalho, Sanjay Awasthi, and Sharda P. Singh
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aging ,sulforaphane ,gut microbiome ,metabolome ,biomarkers ,Biology (General) ,QH301-705.5 - Abstract
Dietary factors modulate interactions between the microbiome, metabolome, and immune system. Sulforaphane (SFN) exerts effects on aging, cancer prevention and reducing insulin resistance. This study investigated effects of SFN on the gut microbiome and metabolome in old mouse model compared with young mice. Young (6–8 weeks) and old (21–22 months) male C57BL/6J mice were provided regular rodent chow ± SFN for 2 months. We collected fecal samples before and after SFN administration and profiled the microbiome and metabolome. Multi-omics datasets were analyzed individually and integrated to investigate the relationship between SFN diet, the gut microbiome, and metabolome. The SFN diet restored the gut microbiome in old mice to mimic that in young mice, enriching bacteria known to be associated with an improved intestinal barrier function and the production of anti-inflammatory compounds. The tricarboxylic acid cycle decreased and amino acid metabolism-related pathways increased. Integration of multi-omic datasets revealed SFN diet-induced metabolite biomarkers in old mice associated principally with the genera, Oscillospira, Ruminococcus, and Allobaculum. Collectively, our results support a hypothesis that SFN diet exerts anti-aging effects in part by influencing the gut microbiome and metabolome. Modulating the gut microbiome by SFN may have the potential to promote healthier aging.
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- 2020
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10. 4287 Extracellular vesicles as biomarkers for early detection of pancreatic cancer
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Charles P Hinzman, Shivani Bansal, Yaoxiang Li, Partha Banerjee, and Amrita Cheema
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Medicine - Abstract
OBJECTIVES/GOALS: Pancreatic ductal adenocarcinoma (PDAC) is projected to become the second leading cause of cancer-related deaths by 2030. Though many other cancers have seen improvements in patient survival rates, patients diagnosed with PDAC have a 5-year survival rate of only ~9%. A major contributor to decreased survival is late-stage diagnosis of the disease. New methods of early detection are urgently needed. Extracellular vesicles (EVs) are secreted from cells of all tissue types into the circulation. EVs play important roles in a variety of diseases. They have shown to promote cancer progression and they are being studied as potential biomarkers for disease diagnosis. The purpose of this study was to perform qualitative and quantitative characterization of small-molecule profiles of EVs derived from various pancreatic cancer (PC) and normal pancreas cell lines, to provide proof-of-concept for evaluating the efficacy of leveraging EVs as potential biomarkers of PDAC. METHODS/STUDY POPULATION: EVs were isolated from the conditioned media of six PC and two normal pancreas cell lines using differential ultracentrifugation with filtration. EV enrichment was validated using quantitative ELISA, immunoblot and transmission electron microscopy. Targeted liquid chromatography coupled to mass spectrometry (LC-MS/MS) and untargeted (UPLC-QTOF-MS) metabolomics were used to analyze the biochemical composition of EVs. RESULTS/ANTICIPATED RESULTS: The biochemical profile of PC EVs was found to be significantly different from the profiles of normal cell EVs. Interestingly, amino acids were downregulated in PC EVs as compared to normal cell EVs. However, PC EVs were enriched in lactate and malate. PC EVs also had significant upregulation in other small molecules such as xanthosine, guanosine diphosphate and nicotinamide. DISCUSSION/SIGNIFICANCE OF IMPACT: Our results indicate that the biochemical characterization of EVs using metabolomics has the potential to yield biomarkers which can delineate cancer cell-derived EVs from normal cell-derived EVs. Further work will test the clinical significance of these findings by similar analyses of plasma of PDAC patients. Furthermore, these profiles may be detectable before progression of the disease to late-stage PDAC, leading to the development of assays for earlier diagnosis in patients.
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- 2020
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11. Tumor suppressor RARRES1- A novel regulator of fatty acid metabolism in epithelial cells.
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Sara Maimouni, Naiem Issa, Selina Cheng, Chokri Ouaari, Amrita Cheema, Deepak Kumar, and Stephen Byers
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Medicine ,Science - Abstract
Retinoic acid receptor responder 1 (RARRES1) is silenced in many cancers and is differentially expressed in metabolism associated diseases, such as hepatic steatosis, hyperinsulinemia and obesity. Here we report a novel function of RARRES1 in metabolic reprogramming of epithelial cells. Using non-targeted LC-MS, we discovered that RARRES1 depletion in epithelial cells caused a global increase in lipid synthesis. RARRES1-depleted cells rewire glucose metabolism by switching from aerobic glycolysis to glucose-dependent de novo lipogenesis (DNL). Treatment with fatty acid synthase (FASN) inhibitor, C75, reversed the effects of RARRES1 depletion. The increased DNL in RARRES1-depleted normal breast and prostate epithelial cells proved advantageous to the cells during starvation, as the increase in fatty acid availability lead to more oxidized fatty acids (FAO), which were used for mitochondrial respiration. Expression of RARRES1 in several common solid tumors is also contextually correlated with expression of fatty acid metabolism genes and fatty acid-regulated transcription factors. Pathway enrichment analysis led us to determine that RARRES1 is regulated by peroxisome proliferating activated receptor (PPAR) signaling. These findings open up a new avenue for metabolic reprogramming and identify RARRES1 as a potential target for cancers and other diseases with impaired fatty acid metabolism.
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- 2018
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12. 3393 Biomarkers of Stroke Recovery Study
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Matthew A. Edwardson, Amrita Cheema, Ming Tan, and Alexander Dromerick
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Medicine - Abstract
OBJECTIVES/SPECIFIC AIMS: There are currently no established blood-based biomarkers of recovery and neural repair following stroke in humans. Such biomarkers would be extremely valuable for aiding in stroke prognosis, timing rehabilitation therapies, and designing drugs to augment natural repair mechanisms. Metabolites, including lipids and amino acids, are engaged in many cellular processes and cross the blood-brain barrier more easily than proteins. Recent advances in liquid chromatography / mass spectrometry (LCMS) allow researchers to obtain a biochemical fingerprint of the metabolites in various biofluids. Thus, metabolite biomarkers of neural repair after brain injury are a promising avenue for future research. Objective: Design and conduct a study to identify metabolite changes in the blood associated with good and poor motor recovery following stroke. METHODS/STUDY POPULATION: We launched the Biomarkers of Stroke Recovery (BIOREC) study, which seeks to enroll 70 participants suffering arm motor impairment following stroke and 35 matched controls. BIOREC is a longitudinal observational study. Fasting blood samples are collected at 5, 15, and 30 days post-stroke, processed, and stored in the Georgetown Lombardi biorepository. Outcome measures, including measures of motor impairment, cognition and language, are assessed at 5, 15, 30, and 90 days post-stroke. The primary outcome measure is the upper extremity Fugl-Meyer score. Control participants are matched for age +/− 1 yr, race, gender, cardiovascular comorbities, and statin use through a computer algorithm that screens the entire MedStar electronic health record (EHR). Control participants provide 2 fasting blood samples one month apart. Once all samples are collected and sent for LCMS analysis, logistic regression analysis will identify potential metabolite biomarkers by comparing participants with good recovery to those with poor recovery as well as stroke participants to controls. RESULTS/ANTICIPATED RESULTS: To date, forty stroke participants have enrolled from 4 acute care hospitals in the Washington, DC metro region and completed all study procedures. Twenty stroke participants either dropped out or were withdrawn due to other medical concerns. Stroke patients ended up at a variety of venues following their acute hospitalization including the acute rehabilitation hospital, skilled nursing facilities, and home. We learned to overcome these logistical challenges by traveling to wherever the patients were sent and notifying medical providers of their study participation. In rare cases we have paid to transport patients from skilled nursing facilities to the clinic, which has reduced dropouts. In addition to the stroke participants, we have enrolled 7 healthy control participants using the EHR screening algorithm. DISCUSSION/SIGNIFICANCE OF IMPACT: Performing a longitudinal study in the early recovery phase following stroke is logistically challenging, but feasible. Difficulty in identifying participants with isolated motor impairment requires added effort to eliminate dropouts. Screening the EHR is an effective method to identify matched controls. Future metabolomics analysis of stored blood samples holds promise to identify biomarkers of stroke recovery and neural repair.
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- 2019
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13. 2016 Plasma microRNA markers of upper limb recovery following human stroke
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Matthew A Edwardson, Xiaogang Zhong, Amrita Cheema, and Alexander Dromerick
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Medicine - Abstract
OBJECTIVES/SPECIFIC AIMS: MicroRNAs are small, non-coding RNAs that control gene expression by inhibiting protein translation. Preclinical studies in rodent stroke models suggest that changes in microRNA expression contribute to neural repair mechanisms. To our knowledge, no one has previously assessed microRNA changes during the recovery phase of human stroke. Our goal was to determine whether patients with significant upper limb recovery following stroke have alteration of neural repair-related microRNA expression when compared to those with poor recovery. METHODS/STUDY POPULATION: Plasma was collected at 19 days post-stroke from 27 participants with mild-moderate upper extremity impairment enrolled in the Critical Periods After Stroke Study. MicroRNA expression was assessed using TaqMan microRNA assays (Thermo Fisher Scientific). Good recovery was defined as ≥6 point change in the Action Research Arm Test (ARAT) score from baseline to 6 months. Bioinformatics analysis compared the plasma microRNA expression profiles of participants with good Versus poor recovery. Candidate biomarkers were identified after correcting for multiple comparisons using a false discovery rate
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- 2018
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14. Serum metabolomic response of myasthenia gravis patients to chronic prednisone treatment.
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Manjistha Sengupta, Amrita Cheema, Henry J Kaminski, Linda L Kusner, and Muscle Study Group
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Medicine ,Science - Abstract
Prednisone is often used for the treatment of autoimmune and inflammatory diseases but they suffer from variable therapeutic responses and significant adverse effects. Serum biological markers that are modulated by chronic corticosteroid use have not been identified. Myasthenia gravis is an autoimmune neuromuscular disorder caused by antibodies directed against proteins present at the post-synaptic surface of neuromuscular junction resulting in weakness. The patients with myasthenia gravis are primarily treated with prednisone. We analyzed the metabolomic profile of serum collected from patients prior to and after 12 weeks of prednisone treatment during a clinical trial. Our aim was to identify metabolites that may be treatment responsive and be evaluated in future studies as potential biomarkers of efficacy or adverse effects. Ultra-performance liquid chromatography coupled with electro-spray quadrupole time of flight mass spectrometry was used to obtain comparative metabolomic and lipidomic profile. Untargeted metabolic profiling of serum showed a clear distinction between pre- and post-treatment groups. Chronic prednisone treatment caused upregulation of membrane associated glycerophospholipids: phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, 1, 2-diacyl-sn glycerol 3 phosphate and 1-Acyl-sn-glycero-3-phosphocholine. Arachidonic acid (AA) and AA derived pro-inflammatory eicosanoids such as 18-carboxy dinor leukotriene B4 and 15 hydroxyeicosatetraenoic acids were reduced. Perturbations in amino acid, carbohydrate, vitamin and lipid metabolism were observed. Chronic prednisone treatment caused increase in membrane associated glycerophospholipids, which may be associated with certain adverse effects. Decrease of AA and AA derived pro-inflammatory eicosanoids demonstrate that immunosuppression by corticosteroid is via suppression of pro-inflammatory pathways. The study identified metabolomic fingerprints that can now be validated as prednisone responsive biomarkers for the improvement in diagnostic accuracy and prediction of therapeutic outcome.
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- 2014
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15. Cholecystokinin Receptor Antagonist Induces Pancreatic Stellate Cell Plasticity Rendering the Tumor Microenvironment Less Oncogenic
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Gurbani Jolly, Tetyana Duka, Narayan Shivapurkar, Wenqiang Chen, Sunil Bansal, Amrita Cheema, and Jill P. Smith
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Cancer Research ,Oncology ,tumor microenvironment ,fibrosis ,pancreatic cancer ,desmoplasia ,CCK ,Tumor microenvironment, fibrosis, pancreatic cancer, Desmoplasia, CC - Abstract
CCK receptors are expressed on pancreatic cancer epithelial cells, and blockade with receptor antagonists decreases tumor growth. Activated pancreatic stellate cells or myofibroblasts have also been described to express CCK receptors, but the contribution of this novel pathway in fibrosis of the pancreatic cancer microenvironment has not been studied. We examined the effects of the nonselective CCK receptor antagonist proglumide on the activation, proliferation, collagen deposition, differential expression of genes, and migration in both murine and human PSCs. CCK receptor expression was examined using western blot analysis. Collagen production using activated PSCs was analyzed by mass spectroscopy and western blot. Migration of activated PSCs was prevented in vitro by proglumide and the CCK-B receptor antagonist, L365,260, but not by the CCK-A receptor antagonist L365,718. Proglumide effectively decreased the expression of extracellular matrix-associated genes and collagen-associated proteins in both mouse and human PSCs. Components of fibrosis, including hydroxyproline and proline levels, were significantly reduced in PSC treated with proglumide compared to controls. CCK peptide stimulated mouse and human PSC proliferation, and this effect was blocked by proglumide. These investigations demonstrate that targeting the CCK-B receptor signaling pathway with proglumide may alter the plasticity of PSC, rendering them more quiescent and leading to a decrease in fibrosis in the pancreatic cancer microenvironment.
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- 2023
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16. Safety and Dosing Study of a Cholecystokinin Receptor Antagonist in Non-alcoholic Steatohepatitis
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Atoosa Rabiee, Martha D. Gay, Narayan Shivapurkar, Hong Cao, Sandeep Nadella, Coleman I. Smith, James H. Lewis, Sunil Bansal, Amrita Cheema, John Kwagyan, and Jill P. Smith
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Pharmacology ,Liver Cirrhosis ,Proglumide ,Liver ,Non-alcoholic Fatty Liver Disease ,Humans ,Pharmacology (medical) ,Receptors, Cholecystokinin ,Cholecystokinin ,Fibrosis - Abstract
High saturated fat diets have been shown to raise blood levels of cholecystokinin (CCK) and induce nonalcoholic steatohepatitis (NASH). CCK receptors are expressed on stellate cells and are responsible for hepatic fibrosis when activated. The purpose of this study was to test the safety and dose of a CCK receptor antagonist, proglumide, in human participants with NASH. An open-label single ascending dose study was conducted in 18 participants with clinical NASH based upon steatosis by liver ultrasound, elevated hepatic transaminases, and a component of the metabolic syndrome. Three separate cohorts (N = 6 each) were treated with oral proglumide for 12 weeks in a sequential ascending fashion with 800 (Cohort 1), 1,200 (Cohort 2), and 1,600 (Cohort 3) mg/day, respectively. Blood hematology, chemistries, proglumide levels, a biomarker panel for fibrosis, and symptom surveys were determined at baseline and every 4 weeks. Abdominal ultrasounds and transient elastography utilizing FibroScan were obtained at baseline and at Week 12. Proglumide was well tolerated at all doses without any serious adverse events. There was no change in body weight from baseline to Week 12. For Cohorts 1, 2, and 3, the median percent change in alanine aminotransferase was 8.42, -5.05, and -22.23 and median percent change in fibrosis score by FibroScan was 8.13, -5.44, and -28.87 (kPa), respectively. Hepatic steatosis as measured by controlled attenuation parameter score significantly decreased with proglumide, (P 0.05). Blood microRNA biomarkers and serum 4-hydroxyproline were consistent with decreased fibrosis at Week 12 compared with baseline. These findings suggest proglumide exhibits anti-inflammatory and anti-fibrotic properties and this compound is well tolerated in participants with NASH.
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- 2022
17. Comparison of amyloid burden in individuals with Down syndrome versus autosomal dominant Alzheimer's disease: a cross-sectional study
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Anna H Boerwinkle, Brian A Gordon, Julie Wisch, Shaney Flores, Rachel L Henson, Omar H Butt, Nicole McKay, Charles D Chen, Tammie L S Benzinger, Anne M Fagan, Benjamin L Handen, Bradley T Christian, Elizabeth Head, Mark Mapstone, Michael S Rafii, Sid O'Bryant, Florence Lai, H Diana Rosas, Joseph H Lee, Wayne Silverman, Adam M Brickman, Jasmeer P Chhatwal, Carlos Cruchaga, Richard J Perrin, Chengjie Xiong, Jason Hassenstab, Eric McDade, Randall J Bateman, Beau M Ances, Howard J Aizenstein, Howard F Andrews, Karen Bell, Rasmus M Birn, Peter Bulova, Amrita Cheema, Kewei Chen, Isabel Clare, Lorraine Clark, Ann D Cohen, John N Constantino, Eric W Doran, Eleanor Feingold, Tatiana M Foroud, Sigan L Hartley, Christy Hom, Lawrence Honig, Milos D Ikonomovic, Sterling C Johnson, Courtney Jordan, M Ilyas Kamboh, David Keator, William E Klunk MD, Julia K Kofler, William C Kreisl, Sharon J Krinsky- McHale, Patrick Lao, Charles Laymon, Ira T Lott, Victoria Lupson, Chester A Mathis, Davneet S Minhas, Neelesh Nadkarni, Deborah Pang, Melissa Petersen, Julie C Price, Margaret Pulsifer, Eric Reiman, Batool Rizvi, Marwan N Sabbagh, Nicole Schupf, Dana L Tudorascu, Rameshwari Tumuluru, Benjamin Tycko, Badri Varadarajan, Desiree A White, Michael A Yassa, Shahid Zaman, Fan Zhang, Sarah Adams, Ricardo Allegri, Aki Araki, Nicolas Barthelemy, Jacob Bechara, Sarah Berman, Courtney Bodge, Susan Brandon, William Brooks, Jared Brosch, Jill Buck, Virginia Buckles, Kathleen Carter, Lisa Cash, Patricio C Mendez, Jasmin Chua, Helena Chui, Laura Courtney, Gregory Day, Chrismary DeLaCruz, Darcy Denner, Anna Diffenbacher, Aylin Dincer, Tamara Donahue, Jane Douglas, Duc Duong, Noelia Egido, Bianca Esposito, Marty Farlow, Becca Feldman, Colleen Fitzpatrick, Nick Fox, Erin Franklin, Nelly Joseph-Mathurin, Hisako Fujii, Samantha Gardener, Bernardino Ghetti, Alison Goate, Sarah Goldberg, Jill Goldman, Alyssa Gonzalez, Susanne Gräber-Sultan, Neill Graff-Radford, Morgan Graham, Julia Gray, Emily Gremminger, Miguel Grilo, Alex Groves, Christian Haass, Lisa Häslerc, Cortaiga Hellm, Elizabeth Herries, Laura Hoechst-Swisher, Anna Hofmann, David Holtzman, Russ Hornbeck, Yakushev Igor, Ryoko Ihara, Takeshi Ikeuchi, Snezana Ikonomovic, Kenji Ishii, Clifford Jack, Gina Jerome, Erik Johnson, Mathias Jucker, Celeste Karch, Stephan Käser, Kensaku Kasuga, Sarah Keefe, William Klunk, Robert Koeppe, Deb Koudelis, Elke Kuder-Buletta, Christoph Laske, Allan Levey, Johannes Levin, Yan Li, Oscar Lopez, Jacob Marsh, Ralph Martins, Neal S Mason, Colin Masters, Kwasi Mawuenyega, Austin McCullough, Arlene Mejia, Estrella Morenas-Rodriguez, John C Morris, James Mountz, Catherine Mummery, Akemi Nagamatsu, Katie Neimeyer, Yoshiki Niimi, James Noble, Joanne Norton, Brigitte Nuscher, Ulricke Obermüller, Antoinette O'Connor, Riddhi Patira, Lingyan Ping, Oliver Preische, Alan Renton, John Ringman, Stephen Salloway, Peter Schofield, Michio Senda, Nicholas T Seyfried, Kristine Shady, Hiroyuki Shimada, Wendy Sigurdson, Jennifer Smith, Lori Smith, Beth Snitz, Hamid Sohrabi, Sochenda Stephens, Kevin Taddei, Sarah Thompson, Jonathan Vöglein, Peter Wang, Qing Wang, Elise Weamer, Jinbin Xu, and Xiong Xu
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Adult ,Cerebral Cortex ,Amyloid beta-Peptides ,Apolipoproteins E ,Cross-Sectional Studies ,Alzheimer Disease ,Positron-Emission Tomography ,Humans ,Neurology (clinical) ,Middle Aged ,Down Syndrome ,Biomarkers ,Aged - Abstract
Important insights into the early pathogenesis of Alzheimer's disease can be provided by studies of autosomal dominant Alzheimer's disease and Down syndrome. However, it is unclear whether the timing and spatial distribution of amyloid accumulation differs between people with autosomal dominant Alzheimer's disease and those with Down syndrome. We aimed to directly compare amyloid changes between these two groups of people.In this cross-sectional study, we included participants (aged ≥25 years) with Down syndrome and sibling controls who had MRI and amyloid PET scans in the first data release (January, 2020) of the Alzheimer's Biomarker Consortium-Down Syndrome (ABC-DS) study. We also included carriers of autosomal dominant Alzheimer's disease genetic mutations and non-carrier familial controls who were within a similar age range to ABC-DS participants (25-73 years) and had MRI and amyloid PET scans at the time of a data freeze (December, 2020) of the Dominantly Inherited Alzheimer Network (DIAN) study. Controls from the two studies were combined into a single group. All DIAN study participants had genetic testing to determine PSEN1, PSEN2, or APP mutation status. APOE genotype was determined from blood samples. CSF samples were collected in a subset of ABC-DS and DIAN participants and the ratio of amyloid β42 (Aβ42) to Aβ40 (Aβ42/40) was measured to evaluate its Spearman's correlation with amyloid PET. Global PET amyloid burden was compared with regards to cognitive status, APOE ɛ4 status, sex, age, and estimated years to symptom onset. We further analysed amyloid PET deposition by autosomal dominant mutation type. We also assessed regional patterns of amyloid accumulation by estimated number of years to symptom onset. Within a subset of participants the relationship between amyloid PET and CSF Aβ42/40 was evaluated.192 individuals with Down syndrome and 33 sibling controls from the ABC-DS study and 265 carriers of autosomal dominant Alzheimer's disease mutations and 169 non-carrier familial controls from the DIAN study were included in our analyses. PET amyloid centiloid and CSF Aβ42/40 were negatively correlated in carriers of autosomal dominant Alzheimer's disease mutations (n=216; r=-0·565; p0·0001) and in people with Down syndrome (n=32; r=-0·801; p0·0001). There was no difference in global PET amyloid burden between asymptomatic people with Down syndrome (mean 18·80 centiloids [SD 28·33]) versus asymptomatic mutation carriers (24·61 centiloids [30·27]; p=0·11) and between symptomatic people with Down syndrome (77·25 centiloids [41·76]) versus symptomatic mutation carriers (69·15 centiloids [51·10]; p=0·34). APOE ɛ4 status and sex had no effect on global amyloid PET deposition. Amyloid deposition was elevated significantly earlier in mutation carriers than in participants with Down syndrome (estimated years to symptom onset -23·0 vs -17·5; p=0·0002). PSEN1 mutations primarily drove this difference. Early amyloid accumulation occurred in striatal and cortical regions for both mutation carriers (n=265) and people with Down syndrome (n=128). Although mutation carriers had widespread amyloid accumulation in all cortical regions, the medial occipital regions were spared in people with Down syndrome.Despite minor differences, amyloid PET changes were similar between people with autosomal dominant Alzheimer's disease versus Down syndrome and strongly supported early amyloid dysregulation in individuals with Down syndrome. Individuals with Down syndrome aged at least 35 years might benefit from early intervention and warrant future inclusion in clinical trials, particularly given the relatively high incidence of Down syndrome.The National Institute on Aging, Riney and Brennan Funds, the Eunice Kennedy Shriver National Institute of Child Health and Human Development, the German Center for Neurodegenerative Diseases, and the Japan Agency for Medical Research and Development.
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- 2022
18. NAD metabolism modulates inflammation and mitochondria function in diabetic kidney disease
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Komuraiah Myakala, Xiaoxin X Wang, Nataliia V. Shults, Bryce A. Jones, Xiaoping Yang, Avi Z Rosenberg, Brandon Ginley, Pinaki Sarder, Leonid Brodsky, Yura Jang, Chan Hyun Na, Yue Qi, Xu Zhang, Udayan Guha, Ci Wu, Shivani Bansal, Junfeng Ma, Amrita Cheema, Chris Albanese, Matthew D Hirschey, Teruhiko Yoshida, Jeffrey B. Kopp, Julia Panov, and Moshe Levi
- Abstract
Diabetes mellitus is the leading cause of cardiovascular and renal disease in the United States. In spite of the beneficial interventions available for patients with diabetes, there remains a need for additional therapeutic targets and therapies in diabetic kidney disease (DKD). Inflammation and oxidative stress are increasingly recognized as important causes of renal diseases. Inflammation is closely associated with mitochondrial damage. The molecular connection between inflammation and mitochondrial metabolism remains to be elucidated. Recently, nicotinamide adenine nucleotide (NAD+) metabolism has been found to regulate immune function and inflammation. In the present studies we tested the hypothesis that enhancing NAD metabolism could prevent inflammation in and progression of DKD. We found that treatment ofdb/dbmice with type 2 diabetes with nicotinamide riboside (NR) prevented several manifestations of kidney dysfunction (i.e., albuminuria, increased urinary kidney injury marker-1 (KIM1) excretion and pathologic changes). These effects were associated with decreased inflammation, at least in part via inhibiting the activation of the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) signaling pathway. An antagonist of the serum stimulator of interferon genes (STING) and whole-body STING deletion in diabetic mice showed similar renoprotection. Further analysis found that NR increased SIRT3 activity and improved mitochondrial function, which led to decreased mitochondrial DNA damage, a trigger for mitochondrial DNA leakage which activates the cGAS-STING pathway. Overall, these data show that NR supplementation boosted NAD metabolism to augment mitochondrial function, reducing inflammation and thereby preventing progression of diabetic kidney disease.
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- 2021
19. Tu1322: PHASE 1 STUDY TO TEST SAFETY AND DOSE OF A CHOLECYSTOKININ RECEPTOR ANTAGONIST, PROGLUMIDE, IN NON-ALCOHOLIC STEATOHEPATITIS (NASH)
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Atoosa Rabiee, Martha D. Gay, Narayan Shivapurkar, Hong Cao, Sandeep Nadella, Coleman Smith, James H. Lewis, Sunil Bansal, Amrita Cheema, John Kwagyan, and Jill P. Smith
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Hepatology ,Gastroenterology - Published
- 2022
20. Correction: Tumor suppressor RARRES1 links tubulin deglutamylation to mitochondrial metabolism and cell survival
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Sara Maimouni, Mi-Hye Lee, You-Me Sung, Michael Hall, Arpita Roy, Chokri Ouaari, Yoo-Seok Hwang, Justin Spivak, Eric Glasgow, Matthew Swift, Jay Patel, Amrita Cheema, Deepak Kumar, and Stephen Byers
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Oncology - Published
- 2021
21. Transcriptome of Rhesus Macaque (Macaca Mulatta) Exposed to Total-body Irradiation
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Yaoxiang Li, Jatinder Singh, Rency Varghese, Yubo Zhang, Oluseyi Fatanmi, Amrita Cheema, and Vijay Singh
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The field of biodosimetry has seen a paradigm shift towards increased use of molecular phenotyping technologies including omics and miRNA, in addition to conventional cytogenetic techniques. Here, we have used a nonhuman primate (NHP) model to study the impact of gamma-irradiation on alterations in blood-based gene expression. We followed eight NHPs for sixty days after exposure to 6.5 Gy gamma-radiation. Analysis of differential gene expression in response to radiation exposure yielded 26,944 dysregulated genes that were not significantly impacted by sex. Further analysis showed an increased association of several pathways including IL-3 Signaling, Ephrin Receptor Signaling, ErbB Signaling, Nitric Oxide Signaling in the cardiovascular system, Wnt/β-catenin signaling, and inflammasome pathway, which were associated with positive survival outcomes in NHPs after acute exposure to radiation. This study provides novel insights into major pathways and networks involved in radiation-induced injuries that may identify biomarkers for radiation injury.
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- 2020
22. Cholecystokinin Receptor Antagonist Improves Efficacy of Chemotherapy in Murine Models of Pancreatic Cancer by Altering the Tumor Microenvironment
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Zoe X. Malchiodi, Hong Cao, Martha D. Gay, Anita Safronenka, Sunil Bansal, Robin D. Tucker, Benjamin A. Weinberg, Amrita Cheema, Narayan Shivapurkar, and Jill P. Smith
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Cancer Research ,Proglumide ,medicine.medical_treatment ,pancreatic cancer ,chemotherapy ,medicine.disease_cause ,Cholecystokinin receptor ,Article ,Fibrosis ,Pancreatic cancer ,CCK receptor ,medicine ,tumor microenvironment ,metastases ,RC254-282 ,Tumor microenvironment ,Chemotherapy ,business.industry ,fibrosis ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,medicine.disease ,Gemcitabine ,Oncology ,Cancer research ,epithelial-to-mesenchymal transition ,Carcinogenesis ,business ,medicine.drug - Abstract
Simple Summary Our research team has identified the cholecystokinin-B receptor (or CCK-BR) as a novel target for treatment of pancreatic cancer. CCK-BRs are over-expressed in pancreatic cancer and activation stimulates growth in cell culture and in animal models. CCK-BRs are also expressed on pancreatic stellate cells—or the fibroblasts that are responsible for the dense fibrosis in the tumor microenvironment that impedes the penetration of chemotherapy. We show strong evidence that treatment with a CCK receptor antagonist, proglumide, decreases fibrosis in the pancreatic tumor microenvironment and increases the influx of T-cells rendering chemotherapy-resistant cancer, sensitive to gemcitabine. Mice treated with the combination therapy had fewer metastases and greater survival. Tumors from these mice had higher gemcitabine levels and novel differentially expressed genes by RNA-Seq. Proglumide is an older drug that was developed many years ago for peptic ulcer disease but is no longer in use. Repurposing this older drug may improve survival from this recalcitrant malignancy. Abstract Pancreatic cancer is resistant to chemotherapy in part due to the dense desmoplastic fibrosis surrounding the tumor, the immunosuppressive cells in the tumor microenvironment (TME), and the early rate of metastases. In this study, we examined the effects of a CCK receptor antagonist, proglumide, alone and in combination with gemcitabine in murine models of pancreatic cancer. Tumor growth rate, metastases, and survival were assessed in mice bearing syngeneic murine or human pancreatic tumors treated with PBS (control), gemcitabine, proglumide, or the combination of gemcitabine and proglumide. Excised tumors were evaluated histologically for fibrosis, immune cells, molecular markers, and uptake of chemotherapy by mass spectroscopy. Peripheral blood was analyzed with a microRNAs biomarker panel associated with fibrosis and oncogenesis. Differentially expressed genes between tumors of mice treated with gemcitabine monotherapy and combination therapy were compared by RNAseq. When given in combination the two compounds exhibited inhibitory effects by decreasing tumor growth rate by 70%, metastases, and prolonging survival. Proglumide monotherapy altered the TME by decreasing fibrosis, increasing intratumoral CD8+ T-cells, and decreasing arginase-positive cells, thus rendering the tumor sensitive to chemotherapy. Proglumide altered the expression of genes involved in fibrosis, epithelial–mesenchymal transition, and invasion. CCK-receptor antagonism with proglumide renders pancreatic cancer susceptible to chemotherapy.
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- 2021
23. Abstract 5326: Cholecystokinin receptor antagonist decreases fibrosis in pancreatic cancer microenvironment to improve uptake and efficacy of chemotherapy
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Zoe X. Malchiodi, Hong Cao, Martha Gay, Sunil Bansal, Benjamin A. Weinberg, Amrita Cheema, Narayan Shivapurkar, and Jill P. Smith
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Cancer Research ,Chemotherapy ,Proglumide ,business.industry ,medicine.medical_treatment ,medicine.disease ,Cholecystokinin receptor ,Gemcitabine ,Oncology ,Fibrosis ,Pancreatic tumor ,Pancreatic cancer ,Cancer research ,Medicine ,business ,medicine.drug ,Cholecystokinin - Abstract
Background: Although pancreatic cancer cells respond to chemotherapeutic agents in cell culture, many of these drugs are less effective in vivo. This decreased response to chemotherapy in animal models and human subjects is in part attributed to the dense stroma of the pancreatic tumor microenvironment (TME) that impedes penetration of drugs. Pancreatic stellate cells and tissue fibroblasts have cholecystokinin (CCK) receptors and when these receptors are activated, they promote collagen deposition and increase fibrosis in the TME. We have previously shown that CCK receptor antagonists can decrease fibrosis and change the T-cell infiltrates of the TME in pancreatic tumors. We hypothesized that an oral CCK receptor antagonist, proglumide, would improve the efficacy of gemcitabine by decreasing fibrosis in the TME and improving uptake of chemotherapy. Methods: 500,000 mT3 murine pancreatic cancer cells were injected subcutaneously into the right flank of 40 C57/BL6 female mice. One week after the mice had measurable tumors (baseline), mice were divided into 4 groups (N=10 mice each) with equal mean tumor volumes. Treatment groups included: PBS control (100µl IP twice weekly); proglumide in drinking water (0.1 mg/ml); gemcitabine (100mg/kg, 100µl IP twice weekly); and combination of gemcitabine and proglumide. Tumors were measured weekly and the volumes were calculated. Mice were euthanized per protocol when the tumor diameter reached 20mm; tumors were removed, then flash frozen for gemcitabine measurement via mass spectrometry. Results: Tumor volumes demonstrated a rapid growth rate in the PBS control mice with a slope of 184.2±2 (mm3/week), whereas growth rates for proglumide (75±2) and gemcitabine monotherapy (72±14) were 2.6-fold slower, and also 3.4-fold slower for mice treated with both gemcitabine and proglumide (55.6±12; p Conclusion: CCK receptor blockade with proglumide decreases fibrosis in pancreatic cancer TME allowing greater tumor uptake of gemcitabine and improving efficacy by slowing tumor growth and improving survival. Strategies to improve first-line therapies for patients with pancreatic cancer may improve overall survival of this disease. Citation Format: Zoe X. Malchiodi, Hong Cao, Martha Gay, Sunil Bansal, Benjamin A. Weinberg, Amrita Cheema, Narayan Shivapurkar, Jill P. Smith. Cholecystokinin receptor antagonist decreases fibrosis in pancreatic cancer microenvironment to improve uptake and efficacy of chemotherapy [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5326.
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- 2020
24. Quantitative Metabolomics
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Amrita Cheema and Siddheshwar Chauthe
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- 2016
25. Abstract 18446: Treatment With Tetrahydrobiopterin Decreases White Matter Injury in a Mouse Model for In Utero Hypoxia in Congenital Heart Disease
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Jennifer Romanowicz, Ludmila Korotcova, Paul Morton, Amrita Cheema, Vittorio Gallo, Richard A Jonas, and Nobuyuki Ishibashi
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Physiology (medical) ,Cardiology and Cardiovascular Medicine - Abstract
Introduction: Reduced oxygen delivery in complex congenital heart disease (CHD) can lead to brain white matter (WM) injury in utero. Currently, no treatment exists. Tetrahydrobiopterin (BH4) is a cofactor for neuronal nitric oxide synthase, and in its absence, toxic peroxynitrite production is favored. Hypoxia activates nitric oxide synthase, which reduces BH4 availability. Hypothesis: Decreased BH4 levels underlie WM injury in the fetus with CHD, and treatment with BH4 will reduce this injury. Methods: Mice were divided into three groups: normoxic controls (Nx), hypoxic (Hx), and hypoxic with BH4 treatment (Hx-BH4). Hx and Hx-BH4 mice were kept at 10.5% FiO2 from postnatal day 3 to 11--a period of WM development equivalent to the 3rd trimester in humans. Brain BH4 levels were quantified and compared between Nx (n=11) and Hx (n=12). Densities of cells expressing CNP (marker of oligodendrocytes--cells responsible for myelination) and Caspase3 (apoptosis marker) were quantified in three WM regions and compared between groups (n=3-6 each). Western blot detected myelin basic protein (myelin marker). Results: Brain BH4 levels were depleted in Hx compared to Nx (-38.4%, p=0.02). CNP+ oligodendrocytes increased after Hx compared to Nx (Fig.1a), consistent with hypoxia-induced proliferation seen previously. BH4 treatment did not limit this proliferation (Fig.1a). Hx had increased WM apoptosis (Fig.1b), which decreased with BH4 treatment (Fig.1b). Remarkably, there was no difference in WM caspase3+ cells between Nx and Hx-BH4 (Fig.1b). There was no difference in effect across WM region. Finally, loss of myelin with hypoxia was mitigated by BH4 treatment (Fig.1c). Conclusions: Our results show that suboptimal BH4 levels influence hypoxic WM injury. BH4 treatment of phenylketonuria is safe during pregnancy, thus maternal BH4 therapy is feasible. Our data demonstrate that repurposing BH4 for use during fetal brain development has potential to limit WM injury in CHD.
- Published
- 2015
26. Enhanced expression of SOS1 is detected in prostate cancer epithelial cells from African-American men
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Olga A, Timofeeva, Xueping, Zhang, Habtom W, Ressom, Rency S, Varghese, Bhaskar V S, Kallakury, Kan, Wang, Youngmi, Ji, Amrita, Cheema, Mira, Jung, Milton L, Brown, Johng S, Rhim, and Anatoly, Dritschilo
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Male ,Time Factors ,Black People ,Adenocarcinoma ,White People ,Article ,Cell Movement ,Risk Factors ,Cell Line, Tumor ,Biomarkers, Tumor ,Humans ,Genetic Predisposition to Disease ,Neoplasm Invasiveness ,RNA, Messenger ,Aged ,Cell Proliferation ,Neoplasm Staging ,Oligonucleotide Array Sequence Analysis ,Mitogen-Activated Protein Kinase 1 ,Prostatectomy ,Mitogen-Activated Protein Kinase 3 ,Reverse Transcriptase Polymerase Chain Reaction ,Gene Expression Profiling ,Prostatic Neoplasms ,Epithelial Cells ,Middle Aged ,Immunohistochemistry ,Up-Regulation ,Gene Expression Regulation, Neoplastic ,Phenotype ,Tissue Array Analysis ,Lymphatic Metastasis ,RNA Interference ,SOS1 Protein - Abstract
African-American (AA) men experience an increased risk of developing prostate cancers as well as increased mortality following treatment as compared to European-American (EA) men. The aim of our study was to identify biological factors with the potential to predispose AA men to prostate tumor progression and metastasis. To identify cancer-specific gene expression patterns in AA men, we established primary prostate cancer epithelial cells from 14 AA and 13 EA men. High-throughput microarrays were used to investigate differences in global gene expression comparing the two groups. Quantitative RT-PCR and immunohistochemistry validated mRNA and protein expression levels. RNAi knockdowns provided support for biological significance for the identified genes in prostate cancer cells. Son of sevenless homolog 1 (SOS1) was overexpressed in AA male-derived primary prostate cancer epithelial cells. Depletion of SOS1 in PC3 and DU145 prostate cancer cells resulted in decreased capacities for cell proliferation, migration and invasion, at least partially through inhibition of extracellular signal-regulated kinase 1 and 2. Tissue microarray analyses of SOS1 expression in prostate carcinomas correlated with Gleason's grades of tumors, consistent with a possible role in prostate cancer progression. Investigation of prostate cancer-derived epithelial cells has led to identification of SOS1 as a potential candidate biomarker and molecular therapeutic target in prostate cancer in AA men, consistent with the hypothesis that a biological basis exists for prostate cancer aggressiveness in AA men.
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- 2009
27. Fluorescent oligonucleotides can serve as suitable alternatives to radiolabeled oligonucleotides
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Rahul, Ballal, Amrita, Cheema, Waaqar, Ahmad, Eliot M, Rosen, and Tapas, Saha
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Radioisotopes ,DNA Repair ,Recombinant Fusion Proteins ,Ubiquitin-Protein Ligases ,Genes, BRCA1 ,Deoxyguanosine ,Nucleic Acid Hybridization ,Breast Neoplasms ,Articles ,Adenocarcinoma ,Transfection ,DNA Glycosylases ,Neoplasm Proteins ,Substrate Specificity ,Oligodeoxyribonucleotides ,8-Hydroxy-2'-Deoxyguanosine ,Cell Line, Tumor ,Isotope Labeling ,Humans ,Female ,Oligonucleotide Probes ,Fluorescein-5-isothiocyanate ,Fluorescent Dyes - Abstract
Prolonged exposure to radiation from radionuclei used in medical research can cause DNA damage and mutation, which lead to several diseases including cancer. Radioactivity-based experiments are expensive and associated with specialized training, dedication of instruments, approvals, and cleanup with potential hazardous waste. The objective of this study was to find an alternative to the use of radioactivity in medical research using nucleic acid chemistry. FITC-labeled oligonucleotides that contain wild-type (wt) and modified base (8-oxo-G) at the same position and their complementary unlabeled strand were synthesized. Purified DNA repair enzyme, OGG1, and nuclear lysates from MCF-7 breast cancer cells were incubated with double-stranded FITC-labeled wt and 8-oxo-G oligonucleotide to demonstrate the OGG1 incision assay. We found that FITC-coupled oligonucleotides do not impose a steric hindrance during duplex formation, and the fluorescence intensity of the oligonucleotide is comparable with the intensity of the radioactive oligonucleotide. Moreover, we have seen that the OGG1 incision assay can be performed using these fluorescence oligonucleotides, replacing conventional use of radiolabeled oligonucleotides in the assay. Although the use of fluorescent-labeled oligonucleotides was described in detail for incision assays, the technique can be applied to replace a broad range of experiments, where radioactive oligonucleotides are used, eliminating the hazardous consequences of radiation.
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- 2009
28. Analysis of LCâMS Data for Characterizing the Metabolic Changes in Response to Radiation.
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Rency S. Varghese, Amrita Cheema, Prabhdeep Cheema, Marc Bourbeau, Leepika Tuli, Bin Zhou, Mira Jung, Anatoly Dritschilo, and Habtom W. Ressom
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- 2010
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29. Evidence of complete cellular repair of 1,N6-ethenoadenine, a mutagenic and potential damage for human cancer, revealed by a novel method.
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Sujata Choudhury, Sanjay Adhikari, Amrita Cheema, and Rabindra Roy
- Abstract
Abstract 1,N6-Ethenoadenine (εA) is generated endogenously by lipid peroxidation and exogenously by tumorigenic industrial agents, vinyl chloride, and vinyl carbamate. εA detected in human tissues causes mutation and is implicated in liver, colon and lung cancers. N-methyl purine DNA-glycosylase (MPG) is the only enzyme known so far to repair εA. However, the mechanism of in vivo repair of εA and the role of MPG remain enigmatic. Moreover, previous in vivo repair studies for DNA lesions, including εA, focused only on the step of the removal of the base lesion without further insight into the completion of the repair process. This may be in part due to the unavailability of an appropriate in vivo quantitative method to evaluate complete BER process at the basal level. Our newly developed in vivo method is highly sensitive and involves phagemid M13mp18, containing εA at a defined position. The complete repair events have been estimated by plaque assay in E. coli with the phagemids recovered from the human cells after cellular processing. We found that the detectable complete (removal and replacement of εA with adenine) repair was observed only 18% in 16 h, but with the repair nearing completion within 24 h in colon cancer, HCT-116, cells. Moreover, MPG is the predominant enzyme for the BER process to remove εA in mammalian cells. Although, the εA is fairly a bulky adduct compared to other small BER substrate lesions, NER pathway is not involved in repair of this adduct. Furthermore, the εA repair in vivo and in vitro is predominant in the G0/G1 phase of the cell cycle. [ABSTRACT FROM AUTHOR]
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- 2008
- Full Text
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30. Small molecule metabolite extraction strategy for improving LC/MS detection of cancer cell metabolome
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Kd, Sheikh, Khanna S, Sw, Byers, Fornace A, and Amrita Cheema
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