This document discusses epigenetic modulation through inhibition of histone demethylases like LSD1. It summarizes that:
1) Polyamino(bis)guanidines and polyaminobiguanides can inhibit the histone demethylase LSD1 in vitro and in human colon cancer cells.
2) These inhibitors are non-competitive inhibitors of LSD1 and promote increased histone H3 lysine 4 dimethylation.
3) One inhibitor, verlindamycin (compound 2d), re-expresses tumor suppressor genes silenced in cancer cells and reduces tumor growth in mouse models of human colon cancer, especially in combination with 5-azacytidine.
Understanding Methylation, Gene Regulation and the MethylDetox ProfileCell Science Systems
This exciting educational presentation shines light on the methylation cycle and discusses the role of nutrients as a clinical strategy. This event will be in a round table format including in-studio discussion surrounding the topics presented. In this round-table, Dino Celeda, Ph.D is accompanied by Amy Pieczarka, RD, LDN, CCN, CDE and Andrew Campbell, M.D.
Key Topics covered during this presentation:
• Learn about methylation and methylation potential
• Discover how methylation potential is correlated with neurotransmitter balance
• Learn the significance of the methionine and homocysteine ratio
• See how the MTHFR gene affects homocysteine balance
• Understand patient methylation with the Methyl Detox Profile
KDM5 epigenetic modifiers as a focus for drug discoveryChristopher Wynder
A summary presentation of my scientific work.
My laboratory focused on an enzyme KDM5b (aka PLU-1, JARID1b) that was widely expressed during development and played a key role in progression of breast cancer through HER-2.
My lab focused on understanding the key biochemical activity of the enzyme through dissecting the proteomic and genomic interactors.
Our results were confirmed through the use of ES cells, adult stem cells and mouse models.
Much of this work remains unpublished, please contact me for more information and/or access to any reagents that I still have as part of this work.
crwynder@gmail.com
Understanding Methylation, Gene Regulation and the MethylDetox ProfileCell Science Systems
This exciting educational presentation shines light on the methylation cycle and discusses the role of nutrients as a clinical strategy. This event will be in a round table format including in-studio discussion surrounding the topics presented. In this round-table, Dino Celeda, Ph.D is accompanied by Amy Pieczarka, RD, LDN, CCN, CDE and Andrew Campbell, M.D.
Key Topics covered during this presentation:
• Learn about methylation and methylation potential
• Discover how methylation potential is correlated with neurotransmitter balance
• Learn the significance of the methionine and homocysteine ratio
• See how the MTHFR gene affects homocysteine balance
• Understand patient methylation with the Methyl Detox Profile
KDM5 epigenetic modifiers as a focus for drug discoveryChristopher Wynder
A summary presentation of my scientific work.
My laboratory focused on an enzyme KDM5b (aka PLU-1, JARID1b) that was widely expressed during development and played a key role in progression of breast cancer through HER-2.
My lab focused on understanding the key biochemical activity of the enzyme through dissecting the proteomic and genomic interactors.
Our results were confirmed through the use of ES cells, adult stem cells and mouse models.
Much of this work remains unpublished, please contact me for more information and/or access to any reagents that I still have as part of this work.
crwynder@gmail.com
Event Details
This webinar will introduce the Advanced MethylDetox Profile, discuss the scientific underpinnings of methylation and detoxifications, and explain how this test can benefit your patients. Our speakers have a diverse range of backgrounds from research to clinical practice.
Key Learning Points
-Discover the critical genes in the methylation pathway
-Understand each gene’s role in patient methylation function
-See how the MethylDetox Profile can be used clinically
-Learn how to monitor treatment progress
PhD poster presented at; the University of Nottingham Pharmacy School 2nd Year PhD Poster Day, the British Association of Cancer Research (BACR) Special Conference: Advances in Cancer Drug Discovery, the University of Nottingham - Faculty of Medicine and Health Sciences Review of Research 2009 and the University of Nottingham Graduate School Poster Day (prize winner). I also presented this poster to the Vitae Midlands Hub Regional Poster Competition 2009 where I represented the University of Nottingham.
Event Details
This webinar will introduce the Advanced MethylDetox Profile, discuss the scientific underpinnings of methylation and detoxifications, and explain how this test can benefit your patients. Our speakers have a diverse range of backgrounds from research to clinical practice.
Key Learning Points
-Discover the critical genes in the methylation pathway
-Understand each gene’s role in patient methylation function
-See how the MethylDetox Profile can be used clinically
-Learn how to monitor treatment progress
PhD poster presented at; the University of Nottingham Pharmacy School 2nd Year PhD Poster Day, the British Association of Cancer Research (BACR) Special Conference: Advances in Cancer Drug Discovery, the University of Nottingham - Faculty of Medicine and Health Sciences Review of Research 2009 and the University of Nottingham Graduate School Poster Day (prize winner). I also presented this poster to the Vitae Midlands Hub Regional Poster Competition 2009 where I represented the University of Nottingham.
The epigenetic regulation of DNA-templated processes has been intensely studied over the last 15
years. DNA methylation, histone modification, nucleosome remodeling, and RNA-mediated targeting regulate many biological processes that are fundamental to the genesis of cancer. Here, we
present the basic principles behind these epigenetic pathways and highlight the evidence suggesting that their misregulation can culminate in cancer. This information, along with the promising clinical and preclinical results seen with epigenetic drugs against chromatin regulators, signifies that it
is time to embrace the central role of epigenetics in cancer.
STAT3, a member of the STAT family, is a latent transcription factor that is activated in response to various cytokines, growth factors, and oncogene signals. STAT3 is constitutively activated in various human cancers, and its activation is frequently associated with poor prognosis. As a transcription factor, STAT3 regulates a set of genes implicated in cancer cell survival, proliferation, angiogenesis, invasion, metastasis, drug resistance, and immune evasion. Medicilon provides STAT3 drug discovery, CMC research (API + formulation), pharmacodynamics research, PK study, safety evaluation and other services.
https://www.medicilon.com/platform/stat3-targeted-drugs/
Green Gasoline and U.S. Energy IndependenceEngenuitySC
In the first installment of our second Year of the SmartState series, Dr. Regalbuto, of the University of South Carolina & the new Energy Leadership Institute (ELI), talks Green Gasoline and U.S Energy Independence in conjunction with Energy Action Month.
Dirk Brown: Pandoodle, Patents, and the Palmetto StateEngenuitySC
Dirk Brown does 3 presentations in one as he shares his story of Pandoodle, an adaptive digital design firm which can places animations into live video, and its journey from Silicon Valley to Columbia, S.C.
Science Cafe Discovers a New Form of Alternative EnergyEngenuitySC
These are the slides from the May Science Cafe featuring Dr. MVS Chandrashekhar. During this cafe he discussed his work with graphene a new, clean energy source.
ISI 2024: Application Form (Extended), Exam Date (Out), EligibilitySciAstra
The Indian Statistical Institute (ISI) has extended its application deadline for 2024 admissions to April 2. Known for its excellence in statistics and related fields, ISI offers a range of programs from Bachelor's to Junior Research Fellowships. The admission test is scheduled for May 12, 2024. Eligibility varies by program, generally requiring a background in Mathematics and English for undergraduate courses and specific degrees for postgraduate and research positions. Application fees are ₹1500 for male general category applicants and ₹1000 for females. Applications are open to Indian and OCI candidates.
The use of Nauplii and metanauplii artemia in aquaculture (brine shrimp).pptxMAGOTI ERNEST
Although Artemia has been known to man for centuries, its use as a food for the culture of larval organisms apparently began only in the 1930s, when several investigators found that it made an excellent food for newly hatched fish larvae (Litvinenko et al., 2023). As aquaculture developed in the 1960s and ‘70s, the use of Artemia also became more widespread, due both to its convenience and to its nutritional value for larval organisms (Arenas-Pardo et al., 2024). The fact that Artemia dormant cysts can be stored for long periods in cans, and then used as an off-the-shelf food requiring only 24 h of incubation makes them the most convenient, least labor-intensive, live food available for aquaculture (Sorgeloos & Roubach, 2021). The nutritional value of Artemia, especially for marine organisms, is not constant, but varies both geographically and temporally. During the last decade, however, both the causes of Artemia nutritional variability and methods to improve poorquality Artemia have been identified (Loufi et al., 2024).
Brine shrimp (Artemia spp.) are used in marine aquaculture worldwide. Annually, more than 2,000 metric tons of dry cysts are used for cultivation of fish, crustacean, and shellfish larva. Brine shrimp are important to aquaculture because newly hatched brine shrimp nauplii (larvae) provide a food source for many fish fry (Mozanzadeh et al., 2021). Culture and harvesting of brine shrimp eggs represents another aspect of the aquaculture industry. Nauplii and metanauplii of Artemia, commonly known as brine shrimp, play a crucial role in aquaculture due to their nutritional value and suitability as live feed for many aquatic species, particularly in larval stages (Sorgeloos & Roubach, 2021).
ANAMOLOUS SECONDARY GROWTH IN DICOT ROOTS.pptxRASHMI M G
Abnormal or anomalous secondary growth in plants. It defines secondary growth as an increase in plant girth due to vascular cambium or cork cambium. Anomalous secondary growth does not follow the normal pattern of a single vascular cambium producing xylem internally and phloem externally.
Toxic effects of heavy metals : Lead and Arsenicsanjana502982
Heavy metals are naturally occuring metallic chemical elements that have relatively high density, and are toxic at even low concentrations. All toxic metals are termed as heavy metals irrespective of their atomic mass and density, eg. arsenic, lead, mercury, cadmium, thallium, chromium, etc.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
Phenomics assisted breeding in crop improvementIshaGoswami9
As the population is increasing and will reach about 9 billion upto 2050. Also due to climate change, it is difficult to meet the food requirement of such a large population. Facing the challenges presented by resource shortages, climate
change, and increasing global population, crop yield and quality need to be improved in a sustainable way over the coming decades. Genetic improvement by breeding is the best way to increase crop productivity. With the rapid progression of functional
genomics, an increasing number of crop genomes have been sequenced and dozens of genes influencing key agronomic traits have been identified. However, current genome sequence information has not been adequately exploited for understanding
the complex characteristics of multiple gene, owing to a lack of crop phenotypic data. Efficient, automatic, and accurate technologies and platforms that can capture phenotypic data that can
be linked to genomics information for crop improvement at all growth stages have become as important as genotyping. Thus,
high-throughput phenotyping has become the major bottleneck restricting crop breeding. Plant phenomics has been defined as the high-throughput, accurate acquisition and analysis of multi-dimensional phenotypes
during crop growing stages at the organism level, including the cell, tissue, organ, individual plant, plot, and field levels. With the rapid development of novel sensors, imaging technology,
and analysis methods, numerous infrastructure platforms have been developed for phenotyping.
Comparing Evolved Extractive Text Summary Scores of Bidirectional Encoder Rep...University of Maribor
Slides from:
11th International Conference on Electrical, Electronics and Computer Engineering (IcETRAN), Niš, 3-6 June 2024
Track: Artificial Intelligence
https://www.etran.rs/2024/en/home-english/
ESR spectroscopy in liquid food and beverages.pptxPRIYANKA PATEL
With increasing population, people need to rely on packaged food stuffs. Packaging of food materials requires the preservation of food. There are various methods for the treatment of food to preserve them and irradiation treatment of food is one of them. It is the most common and the most harmless method for the food preservation as it does not alter the necessary micronutrients of food materials. Although irradiated food doesn’t cause any harm to the human health but still the quality assessment of food is required to provide consumers with necessary information about the food. ESR spectroscopy is the most sophisticated way to investigate the quality of the food and the free radicals induced during the processing of the food. ESR spin trapping technique is useful for the detection of highly unstable radicals in the food. The antioxidant capability of liquid food and beverages in mainly performed by spin trapping technique.
Remote Sensing and Computational, Evolutionary, Supercomputing, and Intellige...University of Maribor
Slides from talk:
Aleš Zamuda: Remote Sensing and Computational, Evolutionary, Supercomputing, and Intelligent Systems.
11th International Conference on Electrical, Electronics and Computer Engineering (IcETRAN), Niš, 3-6 June 2024
Inter-Society Networking Panel GRSS/MTT-S/CIS Panel Session: Promoting Connection and Cooperation
https://www.etran.rs/2024/en/home-english/
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Ana Luísa Pinho
Functional Magnetic Resonance Imaging (fMRI) provides means to characterize brain activations in response to behavior. However, cognitive neuroscience has been limited to group-level effects referring to the performance of specific tasks. To obtain the functional profile of elementary cognitive mechanisms, the combination of brain responses to many tasks is required. Yet, to date, both structural atlases and parcellation-based activations do not fully account for cognitive function and still present several limitations. Further, they do not adapt overall to individual characteristics. In this talk, I will give an account of deep-behavioral phenotyping strategies, namely data-driven methods in large task-fMRI datasets, to optimize functional brain-data collection and improve inference of effects-of-interest related to mental processes. Key to this approach is the employment of fast multi-functional paradigms rich on features that can be well parametrized and, consequently, facilitate the creation of psycho-physiological constructs to be modelled with imaging data. Particular emphasis will be given to music stimuli when studying high-order cognitive mechanisms, due to their ecological nature and quality to enable complex behavior compounded by discrete entities. I will also discuss how deep-behavioral phenotyping and individualized models applied to neuroimaging data can better account for the subject-specific organization of domain-general cognitive systems in the human brain. Finally, the accumulation of functional brain signatures brings the possibility to clarify relationships among tasks and create a univocal link between brain systems and mental functions through: (1) the development of ontologies proposing an organization of cognitive processes; and (2) brain-network taxonomies describing functional specialization. To this end, tools to improve commensurability in cognitive science are necessary, such as public repositories, ontology-based platforms and automated meta-analysis tools. I will thus discuss some brain-atlasing resources currently under development, and their applicability in cognitive as well as clinical neuroscience.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
EngenuitySC's Science Cafe - March with Dr. Patrick Woster
1. Flipping the Switch: Can You
Turn Genes "On" and "Off?”
Craig J. Kutz, 1,2 Steven L. Holshouser,1 Robert A. Casero, Jr.,3 Donald R.
Menick2 and Patrick M. Woster1
1Department of Drug Discovery and Biomedical Sciences, Medical University
of South Carolina, 70 President St., Charleston, SC 29425
2Department of Medicine, Medical University of South Carolina, 141 Ashley
Ave., Charleston, SC 29425
3Sidney Kimmel Comprehensive Cancer Institute, Johns Hopkins School of
Medicine, 1650 Orleans St., Baltimore, MD 21231
2. Lou Hawthorne, the Missyplicity Project and
Genetic Savings and Clone
Lou Hawthorne and Missy Rainbow Copycat
Allie and Copycat Rainbow and Copycat
4. DNA Methylation and Histone Modifications:
Compartmentalization of the genome into domains of
different transcriptional potentials (important for
development and differentiation).
hypoacetylated histones
Dense DNA methylation
Histone H3-K9 methylation
Histone H3-K27 methylation
hyperacetylated histones
Low DNA methylation
Histone H3-K4 methylation
From P. Vertino
5. Epigenetics
• Heritable traits that do not involve changes to
the underlying DNA sequence.
• Epigenetic changes can lead to gene
silencing or gene activation, depending on
the chromatin mark involved.
• Regulated by changes in DNA CpG
methylation and histone protein modification.
6. Control of Gene Expression Through Post-Translational
Modification of Histones
Histone PTMs are coordinated with methylation of
CPG Islands at DNA promoter sites.
LYS
or
ARG
NH2
LYS
or
ARG
HN
Histone
Tail
Histone
Tail
Histone
Tail
Histone
Tail
histone acetyltransferase
histone lysine methyltransferase
protein arginine methyltransferase
histone deacetylase
histone demethylase
Altered
Gene
Expression
Post-translational
Modification
epigenetic writers
epigenetic erasers
epigenetic
readers
7. Components of Cellular Epigenetic Modulation
Via Histone Post Translational Modifications
8. Epigenetic Writers, Erasers and Readers
Epigenetic Writers
Histone acetyltransferases (HATs)
GNAT family (Gcn5, PCAF and ELP3)
p300/CBP family (p300 and cyclic AMP-responsive element
binding protein)
MYST family (Tip60 and MYST 1-4)
Histone lysine methyltransferases
Protein lysine methyltransferases (KMTs) – SET1, SET2,
SUV39, EZH1,EZH2, PRDM, other SET, non-SET
All but non-SET contain SU(VAR)3–9, enhancer-of-Zeste, Trihorax)
Protein arginine methyltransferases (PRMTs)
PRMT Type I, PRMT Type II
Histone lysine phosphorylases (H3 Thr3, Ser10, Thr11, and Ser28)
11. Transcriptional control via histone lysine methylation
Methylation of specific lysine residues on histone tails can lead to either transcriptional activation or
repression
17 lysine residues and 7 arginine residues have been shown to undergo methylation/demethylation
10 lysine methyltransferases and nine arginine methyltransferases are known
Lysine demethylases:
lysine-specific demethylase 1 (LSD1) bound to CoREST complex – specific for
H3K4me1 and H3K4me2 (activating chromatin mark)
lysine-specific demethylase 2 (LSD2) - specific for H3K4me1 and H3K4me2 but not bound
to CoREST or another protein complex
LSD1 bound to androgen receptor – specific for H3K9me1 and H3K9me2 (a deactivating
chromatin mark)
Jumonji (JmjC)-domain containing demethylases
JHDM1A – specific for H3K6me1 and H3K6me2
JHDM2A – H3K9me1 and H3K9me2
Other Jumonji demethylases specific for trimethylated lysines
12. Epigenetics and Cancer
• DNA methylation and histone modifications contribute to aberrant gene
silencing.
• A functional link of aberrant epigenetic gene silencing to the pathophysiology
of cancer has been established.
• Tumor-suppressor genes are frequently inactivated in association with
promoter CpG island methylation.
• Aberrant DNA methylation and histone modifications have been shown to
have potential in risk-assessment, early detection, disease classification
and prognosis prediction in a variety of cancers.
• DNA-methyltransferase inhibitors reactivate functional expression of tumor-
suppressor genes silenced in cancer.
Baylin et al. Nature Reviews Cancer 6, 107–116 ,
2006
13. Klose and Zhang, Nat Rev. Molec. Cell Biol, 2007 8, 307-318
a | The LSD1 reaction mechanism detailing the removal
of a mono-methyl group. LSD1 is proposed to mediate
demethylation of mono- and di-methylated lysine residues
through an amine oxidation reaction using FAD as a
cofactor. Loss of the methyl group from mono-methyl lysine
occurs through an imine intermediate (1), which is
hydrolysed to form formaldehyde by a non-enzymatic
process (2). b | A polypeptide backbone cartoon structure
of LSD1 bound to Co-REST and the cofactor FAD. The
two-lobed amine oxidase (AO) domain is shown in orange
and yellow. The Tower domain is in green and the SWIRM
domain in blue. The Co-REST linker region (pink)
associates with the LSD1 Tower domain and the SANT
domain (red) situated at the top of the Tower domain.
c | Depiction of the potential association of LSD1–Co-REST
with nucleosomal DNA. The bottom half shows a
nucleosome with the core histone octamer in the centre
and the associated DNA double helix in blue. The LSD1–
Co-REST complex modelled onto a nucleosome indicates
that the SANT domain of Co-REST (red) could interact with
nucleosomal DNA, whereas LSD1 targets the histone H3
tail where it protrudes from the DNA gyres (shown by the
arrow). d | LSD1 as part of the Co-REST complexes
contributes to repression of neuronal genes in non-neuronal
cells. LSD1 contributes to repression by removing H3K4
methylation. e | When bound to the androgen receptor (AR),
LSD1 is converted from a transcriptional repressor to an
activator by changing the substrate specificity of LSD1 so
that it catalyses the removal of H3K9 methylation.
16. Polyamino(bis)guanidines and Polyaminobiguanides Inhibit Purified LSD1
0
20000
40000
60000
80000
100000
1a
1c
1f
1d
1e
2a
2e
2f
2c
2b
2d
1b
1g
Untreated
(pmol/mgprotein/min)
rLSD1activity
“You can observe a lot by just watching.”
-Yogi Berra
18. Polyamino(bis)guanidines and Polyaminobiguanides are Non-Competitive Inhibitors of LSD1
-100
-50
0
50
100
150
-0.3 -0.2 -0.1 0 0.1 0.2 0.3
1/H3K4me2 (mM)
1/V
0 mM
0.25 mM
0.5 mM
1 mM
2.5 mM
-100
-50
0
50
100
150
-0.3 -0.2 -0.1 0 0.1 0.2 0.3
1/H3K4me2 (mM)
1/V
0 mM
0.25 mM
0.5 mM
1 mM
2.5 mM
Compound 1c
Compound 2d
21. The Polyaminobiguanide Verlindamycin is a Potent Epigenetic Modulator
-Acts as a non-competitive inhibitor of recombinant LSD1/CoREST (KI = 6.7 mM)
-Promotes a 6.5-fold increase in global H3K4me2 in HCT116 cells in vitro; no change in
methylation levels at H3K9 or H3K27
-Causes significant re-expression of aberrantly silenced tumor suppressor proteins
SFRP1, 4 and 5 and GATA 5.
Huang, Y. et al.: Proc. Nat. Acad. Sci. USA 2007, 104,
8023-8028.
Huang, Y. et al.: Clin. Cancer Res. 2009, 15,
7217-7228
22. In vivo effects of compound 2d in the presence and absence of 5-azacytidine
Verlindamycin (2d) Is Effective In Vivo in Combination with 5-Azacytidine
23. Epigenetics in the Heart
Epigenetics refers to alterations in gene expression independent of
the genetic code.
HDACs have been extensively studied in cardiovascular disease
HDAC inhibitors shown to be cardioprotective in both ischemia
reperfusion injury and heart failure
Recent evidence implies a crosstalk, or even an interdependency,
of HDACs with histone demethylases
Chandrasekaran, S. et al.: Histone deacetylases facilitate sodium/calcium exchanger
up-regulation in adult cardiomyocytes. FASEB J. 2009, 23(11), 3851-3864.
HDAC activity causes
histone methylation
24. Assessment of Drug Effects in the Langendorff Heart Model
Normal rabbit heart Heart after ischemia reperfusion injury
25. Left Ventricular
HCT 116 human colorectal tumor xenograft in Balb/c mice
0 10 20 30 40 50 60 70 80 90 100
0
50
100
150
200
Time(mins)
mmHg Developed Pressure
Vehicle (n=6)
2d (n=5)
No IR (n=1)
ReperfusionNo-Flow
Ischemia
**** *** **
****p<0.0001; ***p<0.001; **p<0.01; *p<0.05
26. In vivo effects of compound 2d in the presence and absence of 5-azacytidine
HCT 116 human colorectal tumor xenograft in Balb/c mice
0 10 20 30 40 50 60 70 80 90 100
0
20
40
60
80
Time(mins)
mmHg End Diastolic Pressure
No IR (n=1)ReperfusionNo-Flow
Ischemia **** ***
****p<0.0001; ***p<0.001; **p<0.01; *p<0.05
Vehicle (n=6)
2d (n=5)
Left Ventricular
30. OH
F
CN
Cl O Cl
CN
O Cl
NH2
S
SN
NC
O
Cl
H
NN
S
NC
NH2H2N
EtOH
microwave
90oC, 10min
ether
microwave,
40oC, 5 min
DMSO, K2CO3
microwave, 190oC
6 min
+
LiAlH4
ether, 0oC, 24 h
CH3
CH3
H3C
O
Cl
H
NH
N
N
N
H2N
X
H
NN
N
NH
H2N
General Structure
Scheme 1
21 22 23
24
266
25
R1
R2
R3
R4
Cl
33. Cellular Effects of C1 and C15 in the Calu6 Lung Adenocarcinoma Cell Line
Vehicle
30µM TCP
1µM 6 10µM 7
1µM 6 10µM 7
DAPI F-Actin H3K4me2
Vehicle
30µM TCP
1µM 6 10µM 6
1µM 7 10µM 7
DAPI F-Actin H3K4me2
34. A B
!
C D
!
E
!
Figure S3. Comparison of the cytotoxicity of compounds 6 and 7 to known agents verlindamycin 2 and TCP in 5 cell lines in vitro using a standard MTS
reduction assay. Panel A: CA46 Burlitt’s Lymphoma cell line; Panel B: PC3 human prostate cancer cell line; Panel C: PANC-1 human pancreatic cancer cell
line; Panel D: MDA-MB-231 estrogen receptor negative breast cancer cell line; Panel E: MCF-10A human breast epithelial cell line. In Panels B and C,
verlindamycin 2 was run at 8 mM as a positive control, while in Panels A, D and E a dose-response curve was generated for 2. Each data point is the average of 3
determinations + standard error.
35. 0 10 20 30 40 50 60 70 80 90 100
0
50
100
150
200
Time(mins)
DevelopedPressure(mmHg) Left Ventricular Developed Pressure
(1-hr pretreatment)
Vehicle (n=6)ReperfusionNo-Flow
Ischemia C1 (n=3)
Verlindamycin (n=3)
No IR (n=2)
36. 0 10 20 30 40 50 60 70 80 90 100
0
20
40
60
80
Time(mins)
EndDiastolicPressure(mmHG) Left Ventricular End Diastolic Pressure
(1-hr pretreatment)
No IR (n=2)
ReperfusionNo-Flow
Ischemia C1 (n=3)
Verlindamycin (n=3)
Vehicle (n=6)
38. Figure 1. LSD1/HDAC/CoREST corepressor complex. LSD1 inhibitors or HDAC
inhibitors (HDACi) can independently re-express silenced promoters through post-
translational histone modifications.
Is the Major Effect of Inhibitors of Chromatin Remodeling Enzymes Mediated
at the Epigenetic Complex?
39. Primary feline cardiomyocytes were treated for 3 h with 5 mM verlindamycin (V), 1 mM C1
or 2 mM C15. The co-repressor HDAC:CoREST:LSD1 complex was initially pulled down with an antibody
for HDAC1 and a Western blot for CoREST was performed. The figure shows that verlindamycin and C1
disrupted the interaction between HDAC1 and CoREST, indicating that LSD1 inhibition may cause
disruption of the entire co-repressor complex.
Additional experiments were performed with a pull-down with LSD1 antibody, showing that C1 and
verlindamycin disrupted LSD1:HDAC1 interaction as well.
Pull-down Experiment for HDAC1/CoREST/LSD1 Complex
Wb: CoREST!
NC ! Veh! 2d! C1! C15!
IP: HDAC1!
IgG Veh V C1 C15
CoREST
Non-specific
40. Acknowledgements
MUSC Johns Hopkins University University of Pretoria
Dr. Donald R. Menick Robert A. Casero Lyn-Marie Birkholtz
Isuru Kumarasinghe Tracey Murray-Stewart Bianca Verlinden
Sun Choi Shannon Nowatarski Jandeli Niemand
Steven Holshouser Valentina Battaglia
Melissa Sokolosky Christina Destefano-Shields Jawarhal Nehru University
Craig Kutz Christin Hanigan Rentala Madhubala
Youxuan Li
Hereward “Cliff” Wimborne
Benefactors
NIH grant RO1 CA149095-01