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Article: Overview of programmed electrical stimulation to assess atrial fibrillation susceptibility in mice.

Murphy, Matthew B / Kannankeril, Prince J / Murray, Katherine T

2023 Volume 14, Page(s) 1149023

Abstract: Atrial fibrillation (AF) is the most common human arrhythmia and is associated with increased risk of stroke, dementia, heart failure, and death. Among several animal models that have been used to investigate the molecular determinants of AF, mouse ... ...

Abstract	Atrial fibrillation (AF) is the most common human arrhythmia and is associated with increased risk of stroke, dementia, heart failure, and death. Among several animal models that have been used to investigate the molecular determinants of AF, mouse models have become the most prevalent due to low cost, ease of genetic manipulation, and similarity to human disease. Programmed electrical stimulation (PES) using intracardiac or transesophageal atrial pacing is used to induce AF as most mouse models do not develop spontaneous AF. However, there is a lack of standardized methodology resulting in numerous PES protocols in the literature that differ with respect to multiple parameters, including pacing protocol and duration, stimulus amplitude, pulse width, and even the definition of AF. Given this complexity, the selection of the appropriate atrial pacing protocol for a specific model has been arbitrary. Herein we review the development of intracardiac and transesophageal PES, including commonly used protocols, selected experimental models, and advantages and disadvantages of both techniques. We also emphasize detection of artifactual AF induction due to unintended parasympathetic stimulation, which should be excluded from results. We recommend that the optimal pacing protocol to elicit an AF phenotype should be individualized to the specific model of genetic or acquired risk factors, with an analysis using several definitions of AF as an endpoint.
Language	English
Publishing date	2023-04-11
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2564217-0
ISSN	1664-042X
ISSN	1664-042X
DOI	10.3389/fphys.2023.1149023
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Article ; Online: Optimization of Transesophageal Atrial Pacing to Assess Atrial Fibrillation Susceptibility in Mice.

Murphy, Matthew B / Kim, Kyungsoo / Kannankeril, Prince J / Murray, Katherine T

Journal of visualized experiments : JoVE

2022 , Issue 184

Abstract: Mouse models of genetic and acquired risk factors for atrial fibrillation (AF) have proven valuable in investigating the molecular determinants of AF. Programmed electrical stimulation can be performed using transesophageal atrial pacing as a survival ... ...

Abstract	Mouse models of genetic and acquired risk factors for atrial fibrillation (AF) have proven valuable in investigating the molecular determinants of AF. Programmed electrical stimulation can be performed using transesophageal atrial pacing as a survival procedure, thus enabling serial testing in the same animal. However, numerous pacing protocols exist, which complicates the reproducibility. The present protocol aims to provide a standardized strategy to develop model-specific experimental parameters to improve reproducibility between studies. Preliminary studies are performed to optimize the experimental methods for the specific model under investigation, including age at the time of the study, sex, and parameters of the pacing protocol (e.g., mode of pacing and definition of AF susceptibility). Importantly, care is taken to avoid high stimulus energies, as this can cause stimulation of the ganglionic plexi with inadvertent parasympathetic activation, manifested by exaggerated atrioventricular (AV) block during pacing and often associated with artifactual AF induction. Animals demonstrating this complication must be excluded from the analysis.
MeSH term(s)	Animals ; Atrial Fibrillation/diagnosis ; Atrial Fibrillation/therapy ; Cardiac Pacing, Artificial/adverse effects ; Cardiac Pacing, Artificial/methods ; Electric Stimulation/adverse effects ; Heart Atria ; Mice ; Reproducibility of Results
Language	English
Publishing date	2022-06-29
Publishing country	United States
Document type	Journal Article ; Video-Audio Media ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/64168
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Article ; Online: Isolevuglandins Promote Mitochondrial Dysfunction and Electrophysiologic Abnormalities in Atrial Cardiomyocytes.

Subati, Tuerdi / Yang, Zhenjiang / Murphy, Matthew B / Stark, Joshua M / Trykall, David Z / Davies, Sean S / Barnett, Joey V / Murray, Katherine T

Cells

2024 Volume 13, Issue 6

Abstract: Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia, yet the cellular and molecular mechanisms underlying the AF substrate remain unclear. Isolevuglandins (IsoLGs) are highly reactive lipid dicarbonyl products that mediate oxidative ... ...

Abstract	Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia, yet the cellular and molecular mechanisms underlying the AF substrate remain unclear. Isolevuglandins (IsoLGs) are highly reactive lipid dicarbonyl products that mediate oxidative stress-related injury. In murine hypertension, the lipid dicarbonyl scavenger 2-hydroxybenzylamine (2-HOBA) reduced IsoLGs and AF susceptibility. We hypothesized that IsoLGs mediate detrimental pathophysiologic effects in atrial cardiomyocytes that promote the AF substrate. Using Seahorse XFp extracellular flux analysis and a luminescence assay, IsoLG exposure suppressed intracellular ATP production in atrial HL-1 cardiomyocytes. IsoLGs caused mitochondrial dysfunction, with reduced mitochondrial membrane potential, increased mitochondrial reactive oxygen species (ROS) with protein carbonylation, and mitochondrial DNA damage. Moreover, they generated cytosolic preamyloid oligomers previously shown to cause similar detrimental effects in atrial cells. In mouse atrial and HL-1 cells, patch clamp experiments demonstrated that IsoLGs rapidly altered action potentials (AP), implying a direct effect independent of oligomer formation by reducing the maximum Phase 0 upstroke slope and shortening AP duration due to ionic current modifications. IsoLG-mediated mitochondrial and electrophysiologic abnormalities were blunted or totally prevented by 2-HOBA. These findings identify IsoLGs as novel mediators of oxidative stress-dependent atrial pathophysiology and support the investigation of dicarbonyl scavengers as a novel therapeutic approach to prevent AF.
MeSH term(s)	Animals ; Mice ; Myocytes, Cardiac/metabolism ; Lipids/chemistry ; Reactive Oxygen Species/metabolism ; Atrial Fibrillation ; Mitochondrial Diseases ; Benzylamines
Chemical Substances	isolevuglandin ; Lipids ; Reactive Oxygen Species ; 2-(aminomethyl)phenol (932-30-9) ; Benzylamines
Language	English
Publishing date	2024-03-09
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells13060483
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Article ; Online: Optimizing transesophageal atrial pacing in mice to detect atrial fibrillation.

Murphy, Matthew B / Kim, Kyungsoo / Kannankeril, Prince J / Subati, Tuerdi / Van Amburg, Joseph C / Barnett, Joey V / Murray, Katherine T

American journal of physiology. Heart and circulatory physiology

2021 Volume 322, Issue 1, Page(s) H36–H43

Abstract: Mice are routinely used to investigate molecular mechanisms underlying the atrial fibrillation (AF) substrate. We sought to optimize transesophageal rapid atrial pacing (RAP) protocols for the detection of AF susceptibility in mouse models. Hypertensive ... ...

Abstract	Mice are routinely used to investigate molecular mechanisms underlying the atrial fibrillation (AF) substrate. We sought to optimize transesophageal rapid atrial pacing (RAP) protocols for the detection of AF susceptibility in mouse models. Hypertensive and control C57Bl/6J mice were subjected to burst RAP at a fixed stimulus amplitude. The role of parasympathetic involvement in pacing-related atrioventricular (AV) block and AF was examined using an intraperitoneal injection of atropine. In a crossover study, burst and decremental RAP at twice diastolic threshold were compared for induction of AV block during pacing. The efficacy of burst and decremental RAP to elicit an AF phenotype was subsequently investigated in mice deficient in the lymphocyte adaptor protein (
MeSH term(s)	Adaptor Proteins, Signal Transducing/genetics ; Animals ; Atrial Fibrillation/genetics ; Atrial Fibrillation/physiopathology ; Echocardiography, Transesophageal/instrumentation ; Echocardiography, Transesophageal/methods ; Echocardiography, Transesophageal/standards ; Heart Rate ; Homeodomain Proteins/genetics ; Male ; Mice ; Mice, Inbred C57BL ; Reproducibility of Results ; Transcription Factors/genetics ; Homeobox Protein PITX2
Chemical Substances	Adaptor Proteins, Signal Transducing ; Homeodomain Proteins ; Lnk protein, mouse ; Transcription Factors
Language	English
Publishing date	2021-11-12
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	603838-4
ISSN	1522-1539 ; 0363-6135
ISSN (online)	1522-1539
ISSN	0363-6135
DOI	10.1152/ajpheart.00434.2021
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Article ; Online: LNK/SH2B3 loss of function increases susceptibility to murine and human atrial fibrillation.

Cardiovascular research

2024

Abstract: Aims: The lymphocyte adaptor protein (LNK) is a negative regulator of cytokine and growth factor signaling. The rs3184504 variant in SH2B3 reduces LNK function and is linked to cardiovascular, inflammatory, and hematologic disorders including stroke. In ...

Abstract	Aims: The lymphocyte adaptor protein (LNK) is a negative regulator of cytokine and growth factor signaling. The rs3184504 variant in SH2B3 reduces LNK function and is linked to cardiovascular, inflammatory, and hematologic disorders including stroke. In mice, deletion of Lnk causes inflammation and oxidative stress. We hypothesized that Lnk-/- mice are susceptible to atrial fibrillation (AF) and that rs3184504 is associated with AF and AF-related stroke in humans. During inflammation, reactive lipid dicarbonyls are a major component of oxidative injury, and we further hypothesized that these mediators are critical drivers of the AF substrate in Lnk-/- mice. Methods and results: Lnk-/- or wild-type (WT) mice were treated with vehicle or 2-hydroxybenzylamine (2-HOBA), a dicarbonyl scavenger, for 3 months. Compared to WT, Lnk-/- mice displayed increased AF duration that was prevented by 2-HOBA. In the Lnk-/- atria, action potentials were prolonged with reduced transient outward K+ current, increased late Na+ current, and reduced peak Na+ current, proarrhythmic effects that were inhibited by 2-HOBA. Mitochondrial dysfunction, especially for complex I, was evident in Lnk-/- atria, while scavenging lipid dicarbonyls prevented this abnormality. Tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were elevated in Lnk-/- plasma and atrial tissue, respectively, both of which caused electrical and bioenergetic remodeling in vitro. Inhibition of soluble TNF-α prevented electrical remodeling and AF susceptibility, while IL-1β inhibition improved mitochondrial respiration but had no effect on AF susceptibility. In a large database of genotyped patients, rs3184504 was associated with AF, as well as AF-related stroke. Conclusions: These findings identify a novel role for LNK in the pathophysiology of AF in both experimental mice and in humans. Moreover, reactive lipid dicarbonyls are critical to the inflammatory AF substrate in Lnk-/- mice and mediate the proarrhythmic effects of pro-inflammatory cytokines, primarily through electrical remodeling.
Language	English
Publishing date	2024-02-20
Publishing country	England
Document type	Journal Article
ZDB-ID	80340-6
ISSN	1755-3245 ; 0008-6363
ISSN (online)	1755-3245
ISSN	0008-6363
DOI	10.1093/cvr/cvae036
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Article ; Online: Autologous bone marrow concentrate intradiscal injection for the treatment of degenerative disc disease with three-year follow-up.

Pettine, Kenneth A / Suzuki, Richard K / Sand, Theodore T / Murphy, Matthew B

International orthopaedics

2017 Volume 41, Issue 10, Page(s) 2097–2103

Abstract: Purpose: The purpose of this study is to assess safety and feasibility of intradiscal bone marrow concentrate (BMC) injections to treat low back discogenic pain as an alternative to surgery with three year minimum follow-up.: Methods: A total of 26 ... ...

Abstract	Purpose: The purpose of this study is to assess safety and feasibility of intradiscal bone marrow concentrate (BMC) injections to treat low back discogenic pain as an alternative to surgery with three year minimum follow-up. Methods: A total of 26 patients suffering from degenerative disc disease and candidates for spinal fusion or total disc replacement surgery were injected with 2 ml autologous BMC into the nucleus pulposus of treated lumbar discs. A sample aliquot of BMC was characterized by flow cytometry and CFU-F assay to determine progenitor cell content. Improvement in pain and disability scores and 12 month post-injection MRI were compared to patient demographics and BMC cellularity. Results: After 36 months, only six patients progressed to surgery. The remaining 20 patients reported average ODI and VAS improvements from 56.7 ± 3.6 and 82.1 ± 2.6 at baseline to 17.5 ± 3.2 and 21.9 ± 4.4 after 36 months, respectively. One year MRI indicated 40% of patients improved one modified Pfirrmann grade and no patient worsened radiographically. Cellular analysis showed an average of 121 million total nucleated cells per ml, average CFU-F of 2713 per ml, and average CD34+ of 1.82 million per ml in the BMC. Patients with greater concentrations of CFU-F (>2000 per ml) and CD34+ cells (>2 million per ml) in BMC tended to have significantly better clinical improvement. Conclusions: There were no adverse events related to marrow aspiration or injection, and this study provides evidence of safety and feasibility of intradiscal BMC therapy. Patient improvement and satisfaction with this surgical alternative supports further study of the therapy.
MeSH term(s)	Adolescent ; Adult ; Autografts ; Bone Marrow Transplantation/adverse effects ; Bone Marrow Transplantation/methods ; Feasibility Studies ; Female ; Follow-Up Studies ; Humans ; Intervertebral Disc Degeneration/therapy ; Low Back Pain/therapy ; Lumbar Vertebrae ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Pain Measurement ; Prospective Studies ; Severity of Illness Index ; Treatment Outcome ; Young Adult
Language	English
Publishing date	2017
Publishing country	Germany
Document type	Journal Article
ZDB-ID	80384-4
ISSN	1432-5195 ; 0341-2695
ISSN (online)	1432-5195
ISSN	0341-2695
DOI	10.1007/s00264-017-3560-9
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Article ; Online: Natriuretic Peptide Oligomers Cause Proarrhythmic Metabolic and Electrophysiological Effects in Atrial Myocytes.

Yang, Zhenjiang / Subati, Tuerdi / Kim, Kyungsoo / Murphy, Matthew B / Dougherty, Owen P / Christopher, Isis L / Van Amburg, Joseph C / Woodall, Kaylen K / Barnett, Joey V / Murray, Katherine T

Circulation. Arrhythmia and electrophysiology

2022 Volume 15, Issue 3, Page(s) e010636

Abstract: Background: With aging, the human atrium invariably develops amyloid composed of ANP (atrial natriuretic peptide) and BNP (B-type natriuretic peptide). Preamyloid oligomers are the primary cytotoxic species in amyloidosis, and they accumulate in the ... ...

Abstract	Background: With aging, the human atrium invariably develops amyloid composed of ANP (atrial natriuretic peptide) and BNP (B-type natriuretic peptide). Preamyloid oligomers are the primary cytotoxic species in amyloidosis, and they accumulate in the atrium during human hypertension and a murine hypertensive model of atrial fibrillation susceptibility. We tested the hypothesis that preamyloid oligomers derived from natriuretic peptides cause cytotoxic and electrophysiological effects in atrial cells that promote arrhythmia susceptibility and that oligomer formation is enhanced for a mutant form of ANP linked to familial atrial fibrillation. Methods: Oligomerization was assessed by Western blot analysis. Bioenergic profiling was performed using the Seahorse platform. Mitochondrial dynamics were investigated with immunostaining and gene expression quantitated using quantitative reverse transcription polymerase chain reaction. Action potentials and ionic currents were recorded using patch-clamp methods and intracellular calcium measured using Fura-2. Results: Oligomer formation was markedly accelerated for mutant ANP (mutANP) compared with WT (wild type) ANP. Oligomers derived from ANP, BNP, and mutANP suppressed mitochondrial function in atrial HL-1 cardiomyocytes, associated with increased superoxide generation and reduced biogenesis, while monomers had no effects. In hypertensive mice, atrial cardiomyocytes displayed reduced action potential duration and maximal dV/dT of phase 0, with an elevated resting membrane potential, compared with normotensive mice. Similar changes were observed when atrial cells were exposed to oligomers. mutANP monomers produced similar electrophysiological effects as mutANP oligomers, likely due to accelerated oligomer formation, while ANP and BNP monomers did not. Oligomers decreased Na Conclusions: These findings provide compelling evidence that natriuretic peptide oligomers are novel mediators of atrial arrhythmia susceptibility. Moreover, the accelerated oligomerization by mutANP supports a role for these mediators in the pathophysiology of this mutation in atrial fibrillation.
MeSH term(s)	Animals ; Atrial Fibrillation/etiology ; Atrial Natriuretic Factor/genetics ; Atrial Natriuretic Factor/metabolism ; Atrial Natriuretic Factor/pharmacology ; Heart Atria ; Mice ; Myocytes, Cardiac/metabolism ; Natriuretic Peptide, Brain/metabolism
Chemical Substances	Natriuretic Peptide, Brain (114471-18-0) ; Atrial Natriuretic Factor (85637-73-6)
Language	English
Publishing date	2022-02-25
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2426129-4
ISSN	1941-3084 ; 1941-3149
ISSN (online)	1941-3084
ISSN	1941-3149
DOI	10.1161/CIRCEP.121.010636
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Article ; Online: Evaluation of the Effectiveness of Autologous Bone Marrow Mesenchymal Stem Cells in the Treatment of Chronic Low Back Pain Due to Severe Lumbar Spinal Degeneration: A 12-Month, Open-Label, Prospective Controlled Trial.

Atluri, Sairam / Murphy, Matthew B / Dragella, Ryan / Herrera, Jessica / Boachie-Adjei, Kwadwo / Bhati, Sachi / Manocha, Vivek / Boddu, Navneet / Yerramsetty, Pavan / Syed, Zaid / Ganjam, Meghana / Jain, Divit / Syed, Zaynab / Grandhi, Nikhil / Manchikanti, Laxmaiah

Pain physician

2022 Volume 25, Issue 2, Page(s) 193–207

Abstract: Background: Regenerative medicine interventions are applied to assist in the repair, and to potentially replace or restore damaged tissue through the use of autologous/allogenic biologics and it continues to expand. The anti-inflammatory, ... ...

Abstract	Background: Regenerative medicine interventions are applied to assist in the repair, and to potentially replace or restore damaged tissue through the use of autologous/allogenic biologics and it continues to expand. The anti-inflammatory, immunomodulatory, and regenerative properties of bone marrow mesenchymal stem cells (BM-MSCs), and investigation into their therapeutic efficacy and safety in patients with severe chronic low back pain, have not been demonstrated in controlled studies. Multiple pain generators have been hypothesized to be responsible in severe spinal degeneration and it is difficult to identify a single pain generator; consequently, resulting in inadequate therapeutic results. Objectives: The study was undertaken to evaluate the effectiveness of autologous bone marrow MSCs in the treatment of chronic low back pain due to severe lumbar spinal degeneration with involvement of multiple structures. Study design: Prospective, open-label, nonrandomized, parallel-controlled, 2-arm exploratory study. Setting: A private, specialized, interventional pain management and regenerative medicine clinic. Methods: The treatment group patients received a one-time bone marrow concentrate injection into spinal structures (i.e., discs, facets, spinal nerves, and sacroiliac joints), along with conventional treatment, whereas, the control group received conventional treatment with nonsteroid anti-inflammatory drugs, over-the-counter drugs, structured exercise programs, physical therapy, spinal injections and opioids, etc., as indicated. Outcomes assessment: Outcomes were assessed utilizing multiple instruments, including the Oswestry Disability Index (ODI), Numeric Rating Scale (NRS-11), EuroQOL 5-Dimensional Questionnaire (EQ-5D-3L), Global Mental Health (GMH), and Global Physical Health (GPH). Multiple outcomes were assessed with primary outcomes being minimal clinically important differences (MCID) in ODI scores between the groups and/or a 2-point reduction in pain scores. In the study group, total nucleated cells, colony forming units-fibroblast, CD34-positive cell numbers and platelets were also recorded, along with post-procedure magnetic resonance imaging changes. Outcomes were assessed at 1, 3, 6, and 12 months. Results: Significant improvement was achieved in functional status measured by ODI, pain relief measured by NRS-11, and other parameters measured by EQ-5D-3L, GMH, and GPH, in the study group relative to the control group at all time periods. The results showed significant improvements at 12-month follow-up with 67% of the patients in the study group achieving MCID utilizing ODI when compared to 8% in the control group. Greater than 2-point pain reduction was seen in 74% of the patients at 3 months, 66% of the patients at 6 months, and 56% of the patients at 12 months. Both MCID and pain relief of 2 points were significantly different compared to the control group. Opioid use decreased in the investigational group, whereas, there was a slight increase in the control group. Age, gender, opioid use, and body mass index did not affect the outcomes in the stem cell group. Limitations: Single center, nonrandomized study. Conclusions: The first available controlled study utilizing BM-MSCs in severe degenerative spinal disease with interventions into multiple structures simultaneously, including disc, facet joints, nerve roots, and sacroiliac joint based on symptomatology, showed promising results.
MeSH term(s)	Analgesics, Opioid ; Humans ; Intervertebral Disc Degeneration/complications ; Intervertebral Disc Degeneration/pathology ; Intervertebral Disc Degeneration/therapy ; Low Back Pain/etiology ; Low Back Pain/pathology ; Low Back Pain/therapy ; Lumbar Vertebrae/pathology ; Mesenchymal Stem Cells ; Prospective Studies ; Treatment Outcome
Chemical Substances	Analgesics, Opioid
Language	English
Publishing date	2022-03-24
Publishing country	United States
Document type	Controlled Clinical Trial ; Journal Article
ZDB-ID	2146393-1
ISSN	2150-1149 ; 1533-3159
ISSN (online)	2150-1149
ISSN	1533-3159
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Article: Biocompatible PLGA-Mesoporous Silicon Microspheres for the Controlled Release of BMP-2 for Bone Augmentation.

Minardi, Silvia / Fernandez-Moure, Joseph S / Fan, Dongmei / Murphy, Matthew B / Yazdi, Iman K / Liu, Xuewu / Weiner, Bradley K / Tasciotti, Ennio

Pharmaceutics

2020 Volume 12, Issue 2

Abstract: Bone morphogenetic protein-2 (BMP-2) has been demonstrated to be one of the most vital osteogenic factors for bone augmentation. However, its uncontrolled administration has been associated with catastrophic side effects, which compromised its clinical ... ...

Abstract	Bone morphogenetic protein-2 (BMP-2) has been demonstrated to be one of the most vital osteogenic factors for bone augmentation. However, its uncontrolled administration has been associated with catastrophic side effects, which compromised its clinical use. To overcome these limitations, we aimed at developing a safer controlled and sustained release of BMP-2, utilizing poly(lactic-
Language	English
Publishing date	2020-02-01
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527217-2
ISSN	1999-4923
ISSN	1999-4923
DOI	10.3390/pharmaceutics12020118
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Article ; Online: Synthesis and evaluation of etoposide and podophyllotoxin analogs against topoisomerase IIα and HCT-116 cells.

Murphy, Matthew B / Kumar, Priyanka / Bradley, Amber M / Barton, Christopher E / Deweese, Joseph E / Mercer, Susan L

Bioorganic & medicinal chemistry

2020 Volume 28, Issue 22, Page(s) 115773

Abstract: Etoposide is a widely-used anticancer agent that targets human type II topoisomerases. Evidence suggests that metabolism of etoposide in myeloid progenitor cells is associated with translocations involved in leukemia development. Previous studies suggest ...

Abstract	Etoposide is a widely-used anticancer agent that targets human type II topoisomerases. Evidence suggests that metabolism of etoposide in myeloid progenitor cells is associated with translocations involved in leukemia development. Previous studies suggest halogenation at the C-2' position of etoposide reduces metabolism. Halogens were introduced into the C-2' position by electrophilic aromatic halogenation onto etoposide (ETOP, 1), podophyllotoxin (PPT, 2), and 4-dimethylepipodophyllotoxin (DMEP, 3), and to bridge the gap of knowledge regarding the activity of these metabolically stable analogs. Five halogenated analogs (6-10) were synthesized. Analogs 8-10 displayed variable ability to inhibit DNA relaxation. Analog 9 was the only analog to show concentration-dependent enhancement of Top2-mediated DNA cleavage. Dose response assay results indicated that 8 and 10 were most effective at decreasing the viability of HCT-116 and A549 cancer cell lines in culture. Flow cytometry with 8 and 10 in HCT-116 cells provide evidence of sub-G1 cell populations indicative of apoptosis. Taken together, these results indicate C-2' halogenation of etoposide and its precursors, although metabolically stable, decreases overall activity relative to etoposide.
MeSH term(s)	A549 Cells ; Antineoplastic Agents/chemical synthesis ; Antineoplastic Agents/chemistry ; Antineoplastic Agents/pharmacology ; Cell Proliferation/drug effects ; Cell Survival/drug effects ; DNA Cleavage ; DNA Topoisomerases, Type II/metabolism ; Dose-Response Relationship, Drug ; Drug Screening Assays, Antitumor ; Etoposide/chemical synthesis ; Etoposide/chemistry ; Etoposide/pharmacology ; HCT116 Cells ; Humans ; Molecular Docking Simulation ; Molecular Structure ; Plasmids/drug effects ; Podophyllotoxin/chemical synthesis ; Podophyllotoxin/chemistry ; Podophyllotoxin/pharmacology ; Structure-Activity Relationship ; Topoisomerase II Inhibitors/chemical synthesis ; Topoisomerase II Inhibitors/chemistry ; Topoisomerase II Inhibitors/pharmacology
Chemical Substances	Antineoplastic Agents ; Topoisomerase II Inhibitors ; Etoposide (6PLQ3CP4P3) ; DNA Topoisomerases, Type II (EC 5.99.1.3) ; Podophyllotoxin (L36H50F353)
Language	English
Publishing date	2020-09-30
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1161284-8
ISSN	1464-3391 ; 0968-0896
ISSN (online)	1464-3391
ISSN	0968-0896
DOI	10.1016/j.bmc.2020.115773
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