A5056787589- Acute Lymphoblastic Leukemia research
- Acute Myeloid Leukemia Research
- Lymphoma Diagnosis and Treatment
- Computational Drug Discovery Methods
- Protein Degradation and Inhibitors
- SARS-CoV-2 and COVID-19 Research
- Histone Deacetylase Inhibitors Research
- RNA modifications and cancer
- Advanced biosensing and bioanalysis techniques
- Epigenetics and DNA Methylation
- Drug Transport and Resistance Mechanisms
- COVID-19 Clinical Research Studies
- Cell Image Analysis Techniques
- Metabolomics and Mass Spectrometry Studies
- Mass Spectrometry Techniques and Applications
- Mosquito-borne diseases and control
- Childhood Cancer Survivors' Quality of Life
- Cancer, Hypoxia, and Metabolism
- CRISPR and Genetic Engineering
- Ubiquitin and proteasome pathways
- Animal testing and alternatives
- Click Chemistry and Applications
- Analytical Chemistry and Chromatography
- Sirtuins and Resveratrol in Medicine
- PARP inhibition in cancer therapy
National Center for Advancing Translational Sciences
2020-2025
National Institutes of Health
2015-2024
National Cancer Institute
2015-2023
Center for Cancer Research
2015-2023
Frederick National Laboratory for Cancer Research
2015-2023
Cornell University
2014-2017
Frederick Community College
2017
Howard Hughes Medical Institute
2016
Bloomsburg University
2014
Poly[adenosine diphosphate (ADP)-ribose] polymerases (PARPs) are a family of enzymes that modulate diverse biological processes through covalent transfer ADP-ribose from the oxidized form nicotinamide adenine dinucleotide (NAD(+)) onto substrate proteins. Here we report robust NAD(+) analog-sensitive approach for PARPs, which allows PARP-specific ADP-ribosylation substrates is suitable subsequent copper-catalyzed azide-alkyne cycloaddition reactions. Using this approach, mapped hundreds...
SARS-CoV-2 is the viral pathogen causing COVID19 global pandemic. Consequently, much research has gone into development of preclinical assays for discovery new or repurposing FDA-approved therapies. Preventing entry a host cell would be an effective antiviral strategy. One mechanism occurs when spike protein on surface binds to ACE2 receptor followed by cleavage at two cut sites ("priming") that causes conformational change allowing and membrane fusion. TMPRSS2 extracellular protease domain...
The National Center for Advancing Translational Sciences (NCATS) has developed an online open science data portal its COVID-19 drug repurposing campaign - named OpenData with the goal of making across a range SARS-CoV-2 related assays available in real-time. cover wide spectrum life cycle, including both viral and human (host) targets. In total, over 10,000 compounds are being tested full concentration-response ranges from multiple annotated small molecule libraries, approved drug,...
Abstract Many compounds with potentially reactive chemical motifs and poor physicochemical properties are published as selective modulators of biomolecules without sufficient validation then propagated in the scientific literature useful probes. Several histone acetyltransferase (HAT) inhibitors these liabilities now routinely used to probe epigenetic pathways. We profile most commonly HAT confirm that majority them nonselective interference compounds. Most (15 out 23, 65%) flagged by ALARM...
Robust, generalizable approaches to identify compounds efficiently with undesirable mechanisms of action in complex cellular assays remain elusive. Such a process would be useful for hit triage during high-throughput screening and, ultimately, predictive toxicology drug development. Here we generate cell painting and health profiles 218 prototypical cytotoxic nuisance U-2 OS cells concentration-response format. A diversity that cause damage produces bioactive morphologies, including...
The human acetyltransferase NAT10 has recently been shown to catalyze formation of N4-acetylcytidine (ac4C), a minor nucleobase known alter RNA structure and function. In order better understand the role acetyltransferases in biology disease, here we report development application chemical methods study ac4C. First, demonstrate that ac4C can be conjugated carrier proteins using optimized protocols. Next, describe access ac4C-containing RNAs, enabling screening anti-ac4C antibodies. Finally,...
C646 inhibits the lysine acetyltransferases (KATs) p300 and CBP represents most potent selective small molecule KAT inhibitor identified to date. To gain insights into cellular activity of this epigenetic probe, we applied chemoproteomics identify covalent targets chemotype. Modeling synthetic derivatization was used develop a clickable analogue (C646-yne) that similarly parent compound enables enrichment bound proteins. LC–MS/MS major C646-yne as highly abundant cysteine-containing...
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has prompted researchers to pivot their efforts finding antiviral compounds and vaccines. In this study, we focused on the human host cell transmembrane protease serine (TMPRSS2), which plays an important role in viral life cycle by cleaving spike protein initiate membrane fusion. TMPRSS2 is attractive target received attention for development of drugs against SARS Middle East syndrome. Starting with comparative...
Nicotinamide adenine dinucleotide (NAD) is increasingly recognized as an important signaling molecule that affects numerous biological pathways. Thus, enzymes metabolize NAD can have functions. One NAD-metabolizing enzyme in mammals CD38, a type II transmembrane protein converts primarily to adenosine diphosphate ribose (ADPR) and small amount of cyclic (cADPR). Localization CD38 was originally thought be only on the plasma membrane, but later reports showed either significant or solely,...
Abstract Mutations in epigenetic regulators are common relapsed pediatric acute lymphoblastic leukemia (ALL). Here, we uncovered the mechanism underlying relapse of ALL driven by an activating mutation NSD2 histone methyltransferase (p.E1099K). Using high-throughput drug screening, found that NSD2-mutant cells were specifically resistant to glucocorticoids. Correction this restored glucocorticoid sensitivity. The transcriptional response glucocorticoids was blocked due depressed receptor...
Adult liver malignancies, including intrahepatic cholangiocarcinoma and hepatocellular carcinoma, are the second leading cause of cancer-related deaths worldwide. Most individuals treated with either combination chemotherapy or immunotherapy, respectively, without specific biomarkers for selection. Here using high-throughput screens, proteomics in vitro resistance models, we identify small molecule YC-1 as selectively active against a defined subset cell lines derived from both cancer types....
Abstract Nicotinamide N-methyltransferase (NNMT) has emerged as a cancer therapeutic target master metabolic regulator that catalyzes the differentiation of normal fibroblasts to associated (CAFs) in stromal tissue surrounding various tumors, most notably ovarian carcinomas. NNMT activity depletes S-adenosyl methionine (SAM) reducing histone methylation. This change epigenome results changes interstitial tumor space which turn promotes growth and metastasis We will present development novel...
Remodelin is a putative small molecule inhibitor of the RNA acetyltransferase NAT10 which has shown preclinical efficacy in models premature aging disease Hutchinson–Gilford Progeria Syndrome (HGPS). Here we evaluate remodelin's assay interference characteristics and effects on NAT10-catalyzed cytidine acetylation. We find remodelin chemotype constitutes cryptic compound, does not react with thiols but demonstrates protein reactivity ALARM NMR proteome-wide affinity profiling assays....
The current landscape of small-molecule drug discovery prioritizes genetic dependencies considered as viable protein targets. While this approach has driven significant therapeutic advancements, it also constrained the exploration non-dependency targets harboring tractable vulnerabilities. Moreover, many remain undruggable, precluding their exploitation and leaving critical gaps in addressing cancers with unmet clinical needs. To overcome these limitations, we developed a tandem screening...
Lysine acetyltransferases (KATs) play a critical role in the regulation of transcription and other genomic functions. However, persistent challenge is development assays capable defining KAT activity directly living cells. Toward this goal, here we report application previously reported dCas9-p300 fusion as transcriptional reporter activity. First, benchmark relative to dCas9-based activators demonstrate its compatibility with second generation short guide RNA architectures. Next, repurpose...
SARS-CoV-2 is the viral pathogen causing COVID19 global pandemic. Consequently, much research has gone into development of pre-clinical assays for discovery new or repurposing FDA-approved therapies. Preventing entry a host cell would be an effective antiviral strategy. One mechanism occurs when spike protein on surface binds to ACE2 receptor followed by cleavage at two cut sites ("priming") that causes conformational change allowing and membrane fusion. TMPRSS2 extracellular protease domain...
SARS-CoV-2, the cause of COVID-19 pandemic, exploits host proteins for viral entry into human lung cells and is blocked by otamixaban in combination with a covalent protease inhibitor.
The androgen receptor is a key regulator of prostate cancer and the principal target current therapies collectively termed deprivation therapies. Insensitivity to these drugs hallmark progression terminal disease state castration-resistant cancer. Therefore, novel therapeutic options that slow combine effectively with existing agents are in urgent need. We show JG-98, an allosteric inhibitor HSP70, re-sensitizes by targeting mitochondrial HSP70 (HSPA9) suppress aerobic respiration. Rather...
An approach is described for high-throughput quality assessment of drug candidate libraries using acoustic ejection high-resolution mass spectrometry (AEMS). Sample introduction from 1536-well plates demonstrated this application 2.5 nL acoustically dispensed sample droplets into an Open Port Interface (OPI) with pneumatically assisted electrospray ionization at a rate one second per sample. Both positive and negative are shown to be essential extend the compound coverage protease...
Lysine acetyltransferases (KATs) are critical regulators of signaling in many diseases, including cancer. A major challenge establishing the targetable functions KATs disease is a lack well-characterized, cell-active KAT inhibitors. To confront this challenge, here we report microfluidic mobility shift platform for discovery and characterization small molecule Novel fluorescent peptide substrates were developed four well-known enzymes (p300, Crebbp, Morf, Gcn5). Enzyme-catalyzed acetylation...
Methotrexate (MTX) is a tight-binding dihydrofolate reductase (DHFR) inhibitor, used as both an antineoplastic and immunosuppressant therapeutic. MTX, like folate undergoes folylpolyglutamate synthetase-mediated γ-glutamylation, which affects cellular retention target specificity. Mechanisms of MTX resistance in cancers include decrease poly-γ-glutamylation upregulation DHFR. Here, we report series potent MTX-based proteolysis targeting chimeras (PROTACs) to investigate DHFR degradation...