Two projects. Innate immunity, human gene6cs and resistance to infec6on. HAMLET; a new type of tumor- killing molecular complex
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1 Two projects Innate immunity, human gene6cs and resistance to infec6on HAMLET; a new type of tumor- killing molecular complex
2 Innate immune signaling Toxins d Capsule Metabolic genes Metalbinding proteins TcpC Activation of innate immunity PMNs Dendritic cells Inhibition of innate immunity No/weak innate immunity Chemokines Cytokines Mucosal mast cells Monocytes Lymphocytes Symptomatic infection Asymptomatic carriage
3 P-fimbriated E. coli GSLs TLR4 Type 1-fimbriated E. coli MGPs TLR4 Phenotype in knock-out mice CD14 negative cell membrane r-ceramide MAP2K3 P TRAM * * TRIF PLC γ * * * * P MAP3K1 Pyk2 asymptomatic bacterial carriage Fyn ERK1/2 PKA P JNK RSK1/2/3 p38 Nucleus P CREB P CREB? IRF3/7 P IRF3/7 P * * P * IFNβ Jun Fos P AP1 RANTES NF-κB IκB NF-κB IL-8 IL-6 acute pyelonephritis, bacteremia, renal scarring
4 Molecular basis of acute pyelonephri6s Ines Ambite
5 Why do mice and pa.ents develop severe kidney infec.on and.ssue pathology? Pathogen-specific signalling pathway in kidney epithelial cells: Single gene deletion induced pathology in the mice model: C57Bl/6 GSLs r-ceramide Irf7 -/- Irf3 -/- Ifnb -/-
6 Mouse gene expression analysis: Good versus Bad inflamma.on Pathogen-specific regulated genes in severely damaged kidneys: Cellular relevance on target genes in epithelial cells: IRF7 TLR4 IFNB Medium CFT073 IRF7 TLR4 IL6 Gene expression value *** ***
7 Molecular basis of acute cys66s Karoly Nagy
8 Molecular basis of acute cys66s
9 Virulence genomics Nanoparticles biomarkers diagnostics Nano-particle Innate immune signaling Proteins Chemokines Cytokines Phosphorylation Gene expression Proteomics Biomarkers identification Gene knock-out animals Experimental UTI models Verification in animal model Human inoculation model ABU protections for recurrent UTI Verification in human model DMSA Children with first UTI Children with UTI and high risk to renal scaring UTI prone adults patients Recurrent UTI Urosepsis prone patient Prospective clinical studies
10 Suppression of tumor cell prolifera6on by Escherichia coli Gustav Rydström
11 Escherichia coli ABU-strain Therapeutic ABU 50% of ABU strains evolved from Uropathogenic Escherichia coli (UPEC) Modulation of host gene expression via inhibition of RNA polymerase II activity Anti-tumorigenic effects? Loss of Virulence UPEC E. coli 83972
12 Inhibition of RNA polymerase II activity Downregulation of Ki-67 in stimulated cells POL II B-actin E. coli Control E. coli (10 8 ) Control Total number of cells (10 5 ) Inhibition of proliferation Control E. coli Hours of incubation Percentage of control Decreased BrdU-incorporation P < 0,05 P < 0,05 Control 10^7 10^8
13 HAMLET! Human alpha-lactalbumin made lethal to tumor cells! α-lactalbumin! oleic acid" C18:1:9cis! HAMLET! Ca 2+! Håkansson, PNAS 1995, Svensson, PNAS, 2000, Petterson-Kastberg, J Mol Biol. 2009
14 Small Angle X-ray Scattering (SAXS) " structure of HAMLET! Ho et al. (2012) PLoS ONE 7(12): e53051! Ho et al. (2012) PLoS ONE 7(12): e53051!
15 HAMLET and colon cancer Manoj Puthia
16 Why is HAMLET suitable to treat colon cancer Photo: Davies et al. Nature reviews cancer 2005, 5:199
17 Peroral HAMLET treatment of developed tumors Mouse model: APC min/+ mice carry dominant heterozygous nonsense mutation in APC gene genetically relevant model mimic human intestinal carcinogenesis Develop multiple polyps at 8-10 weeks age 10 days 5 weeks 24 weeks 10 week old APC Min/+ mice HAMLET (10mg orally, twice daily) Sacrifice treatment group Sacrifice survival group 0 h Off-feed Off-water 5h 5.30 h Feed water HAMLET Puthia et al. Gut (in press) 10h Off-feed Off-water 15h h Feed water HAMLET Tumor count Methylene blue staining Microarray Histology IHC
18 HAMLET treatment reduces tumor burden Control Control HAMLET treated HAMLET treated Control HAMLET treated Puthia et al. Gut (2013)
19 HAMLET treatment reduces tumor burden and improves survival in APC min mice Polyp number/ mouse 80 Control HAMLET treated P < P < P > 0.05 P < < >2 Total Polyp diameter (mm) Percent survival P = HAMLET treated Control Weeks Puthia et al. Gut (2013)
20 Welcome to our lab!!!! BMC C12
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