Trent W Nichols - PowerPoint Presentation

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Trent W Nichols

Hepatitis-2015Orlando, USAJuly 20 - 22 2015Slide2

GENE UP AND DOWN-REGULATION WITH MODERATE MAGNETIC FIELDS VIA TOLL-RECEPTORS AND WNT5a; FATTY LIVER/NASH Cirrhosis AND HCC in “MICE AND MEN.”TRENT W. NICHOLS Jr. AMRI, CNDD Hanover PA, VHA Martinsburg WVKEVIN YAREMA. Translational Tissue Engineering Center, Department of Biomedical Engineering Johns Hopkins UniversityBaltimore MD twnicholpa@comcast.netSlide3

GENE UP AND DOWN-REGULATION WITH MODERATE MAGNETIC FIELDS VIA TOLL-RECEPTORS AND WNT5a; FATTY LIVER/NASH Cirrhosis AND HCC in “MICE AND MEN.”Acknowledgements; Funding was provided by Arnold and Mabel Beckman Foundation with the analysis of IL-6-mediated and ganglioside-associated responses funded the NIH (NIBIB, grant 5R01EB5692). The hEDB LVEC cell line y donated by Michael Shamblott (JHMI). Helpful discussions was with Wayne Bonlie and for the gift of the magnetic treatment device from AMRI International (Calgary, AB).The following people have nothing to disclose; Trent W. Nichols Jr., Kevin J. YaremaSlide4

GENE UP AND DOWN-REGULATION WITH MODERATE MAGNETIC FIELDS VIA TOLL-RECEPTORS AND WNT5a; FATTY LIVER/NASH Cirrhosis AND HCC in “MICE AND MEN.”A previous report by L Bi has shown that WNT5a has been shown to be involved in cancer progression and was down regulated in hepatocellular cancer (Hepatology 2013). We have previous reported that many genes in obesity and fatty liver (NAFLD) are associated with insulin resistance and their association to (NASH) non- alcoholic steatohepatitis and the progression to cirrhosis (OMICS J Liver 2013) We now report that many of these genes seen in NASH are regulated by static magnetic fields and that WNT5a is up-regulated and its relevance to oncogenesis.Slide5

Wnt Signaling pathwayBACKGROUND The Wnt signaling pathways are a group of signal transduction pathways made of proteins that pass signals from outside of a cell through cell surface receptors to the inside of the cell. Three Wnt signaling pathways have been characterized: the canonical Wnt pathway, the noncanonical planar cell polarity pathway, and the

noncanonical Wnt/calcium pathway. All three Wnt signaling pathways are activated by the binding of a Wnt-protein ligand to a Frizzled family receptor, which passes the biological signal to the protein Dishevelled inside the cell. The canonical Wnt pathway leads to regulation of gene transcription, the noncanonical planar cell polarity pathway regulates the cytoskeleton that is responsible for the shape of the cell, and the noncanonical Wnt/calcium pathway regulates calcium inside the cell. Slide6

Wnt PathwayWnt signaling pathways use either nearby cell-cell communication (paracrine) or same-cell communication (autocrine). They are highly evolutionarily conserved in animals, which means they are similar across many species of animal from fruit flies to humans.[1][2]Wnt signaling was first identified for its role in carcinogenesis, but has since been recognized for its function in embryonic development. The embryonic processes it controls include body axis patterning, cell fate specification, cell proliferation, and cell migration. Slide7

Wnt Signaling pathwayFigure 1. Wnt doesn't bind to the receptor. Axin, GSK and APC form a "destruction complex," and β-Cat is destroyed.Slide8

Wnt Signaling MechanismWnt signaling begins when one of the Wnt proteins binds to the N-terminal extra-cellular cysteine-rich domain of a Frizzled (Fz) family receptor.[10] These receptors span the plasma membrane seven times and constitute a distinct family of G-protein coupled receptors (GPCRs).[11] However, to facilitate Wnt signaling, co-receptors may also be required alongside the interaction between the Wnt protein and Fz receptor. Examples include lipoprotein receptor-related protein (LRP)-5/6, receptor tyrosine kinase (Ryk), and ROR2.[4] Slide9

Wnt MechanismFigure 2. Wnt binds to (activates) the receptor. Axin is removed from the "destruction complex." β-Cat moves into the nucleus, binds to a transcription factor on DNA, and activates transcription of a protein. "P" represents phosphate.Slide10

BackgroundA previous report by L Bi has shown that WNT5a has been shown to be involved in cancer progression and was down regulated in hepatocellular cancer (Hepatology 2013). We have previous reported that many genes in obesity and fatty liver (NAFLD) are associated with insulin resistance and their association to (NASH) non- alcoholic

steatohepatitis and the progression to cirrhosis. (OMICS J Liver 2013) We now report that many of these genes seen in NASH are regulated by static magnetic fields and that WNT5a is up-regulated .Slide11

Wnt 5a with SMF Signaling METHODOLOGY: Gene array analysis of 2 human embryonic stem cells lines in another experiment of 0.23-0.28 T static magnetic fields (SMF) at JHU as previously described conducted to exam the mechanism of DC EMF via the impact of altered Ca2+ flux and lipid membrane domain fluidity on signaling pathways via TRL4 and IL6. (Wang BMC Genomics 2009). Some 2500 genes were found to up and down-regulated. Recently, further inspection was conducted on Cell Cycle, Cancer, Cell to Cell Signaling, and Cellular Development. Slide12

Ingenuity Network Metabolic Disease, Cellular Development, Connective Tissue Development & FunctionSlide13

Wnt 5a Upregulation by SMFAnalysis of Ingenuity pathways of the Affymetrix gene arrays revealed WNT5a up regulation by SMF after down-regulation of IL6 mRNA at 5 days in stem cell lines. Down-regulation of tumor necrosis factor alpha and up-regulation of TLR 4 were demonstrated for this transformationSlide14

RESULTS: Chemokine genes CCL2, CXCl1, CXCL2, MMP9, SERPINE1,IL6, and IL1B,( genes for fibrosis) were down-regulated. TIMP1, and SOCS3 metalloproteinase inhibitors were up-regulated. Forkhead FOXO1and 3 transcription factors are also up-regulated at 5 days. FOXM1expression when silenced has been found to suppress human HCC cell growth in vitro by Park (J Clin Oncol 2013) Of the 47 genes up-regulated in NASH by a study by Bertola in obesity and NASH (PlosOne 2010), 11 were identified as down-regulated by SMF and 2 up-regulated can prevent fibrosis (See TABLE below)Slide15

TABLE OF GENES DESCRIBED INITAILLY BY Bertola (PLoS One201) FOUND UP and DOWN- REGULATED BY SMF in human stem cells (Wang and Yarema).( SO is no steatosis, S3 steatosis, FOLD N is the decrease or increase in regulation of gene by SMF).

GENE SymbolCONTROLSSO

S3NASHFOLD NASH VS S3 SMFCD54/ICAM11.0060.161.7660.241.88606.4861.10-3.5 - 1.415CXCR1/IR8RA1.0060.182.5460.862.7760.527.3961.31-2.7 -1.509       CXCR2/IL8RB1.0060.211.6660.421.9360.306.1260.973.2 -2.526

CXCL3/GROc1.0060.362.9160.765.0261.2113.8263.59-2.8 -1.311CCL2/MCP11.0060.322.5860.4813.8263.5912.5265.78-4.1 -2.148IL61.0060.424.0160.963.5861.6712.5265.78-5.9 -3.250IL1B1.0060.420.9060.171.4160.277.6862.04-5.4 -1.412TNF1.0060.370.9960.22

1.2860.305.9060.91-4.6 -1.410TNFRSF1B1.0060.261.5460.101.4460.173.4060.85-2.4 -1.295MMP91.0060.292.3560.413.6360.6816.5464.00-4.6 -3.954SERPINE1/PAII1.0060.351.9560.835.6560.7525.3964.18-4.5 -1.477TIMP11.0060.120.9460.191.0460.152.9860.86+2.9 -1.372

SOCS3SOCS11.0060.34 1.0060.113.6360.73 2.0660.443.6460.81 1.7860.2313.3763.34 5.1061.44+3.7 +1.433 2.9 +1.537      -1.351Slide16

Results: : WNT5a up- regulation by SMF leads to NLK down regulation. Nemo Like Kinase -SETDB1 complex subsequently interacts with PPARG. Methylation of PPARG target promotors at histone H3KP and transcriptional silencing inhibiting adipogenesis as been shown by Okamura (Organogenesis 2010). Slide17

CONCLUSIONS: Polygene up regulation and down regulation occurs in fatty liver/NASH. Moderate magnetic field therapy was found in 1 week of therapy of an animal model to reduce fat in Ob/Ob mice (Medical Hypotheses 2012) by this author and in human stem cell lines suggesting that such therapy may be helpful in therapy for NASH to prevent HCC.. Slide18

Conclusions:Adding electrons to the mitochondria Electron Transport Chain is one of the suggested mechanisms. Accelerated liver detoxification by moderate magnetic therapy of obesogens in human subjects that disrupt homeostasis of metabolism of lipids ultimately resulting in obesity is also possible. Activation of AMPK inhibits cancer cell growth through AKT/FOXO3a/FOXM1signaling cascade (Yung BMC Cancer 2013). Polygenic up and down regulation in NASH and its progression to HCC via DC EMF is therefore a possible solution to this epidemic problem.Slide19

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