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Session 1.3: Klampfer
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Session 1.3: Klampfer

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Lidija Klampfer

Lidija Klampfer

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Session 1.3: Klampfer Presentation Transcript

  • 1. Tumor stroma: an active player in cancer
  • 2. Soluble factors produced by stromal cells alter signaling pathways in tumor cells
    HCT116-
    NFkB-LUC
    HCT116-
    Hes1/5-LUC
    Macrophages
    0.4µm
    HCT116-
    TOP-LUC
    HCT116-
    STAT1-LUC
    HCT116-
    STAT3-LUC
    HCT116-
    AP1-LUC
    Tumor cells
  • 3. Colon cancer cells stimulate macrophages to release IL1b;
    the role of pSTAT1
    ( siRNA)
    STAT1
    NSP
    pSTAT1
    STAT3
    -actin
    (IL-1b)
    0 8h 24h 48h
    pGSK3
    c-myc
    tGSK3b
  • 4. Vitamin D3 inhibits the protumorigenic activity of macrophages
    THP1/IL1 -/-
    THP1/D3
    CTRL
    THP1
    pGSK3
    c-myc
    tGSK3b
  • 5. Vitamin D3: an inhibitor of IL1b release from macrophages
    24h
    48h
    1, 25(OH)2D3 (nM)
    50
    50
    0
    0
    25
    25
    pSTAT1
    STAT1
    -actin
    CTRL
    1, 25(OH)2D3 (nM)
  • 6. IL1b mediates the crosstalk between tumor cells
    and macrophages
    ??
    Wnt targets
    b-catenin
    TCF
    STAT1
    pSTAT1
    GSK3b
    NFkB
    AKT
    PDK1
    IL-1b
    Proliferation
  • 7. Macrophages interfere with TRAIL-induced apoptosis
    Hke3
    HCT116
    Trail-Mo
    Trail –IL1
    Trail -Mo
    Trail-Mo
    CTRL
    Trail
    CTRL
    Trail
    Cl caspase 8 (41,43 kDa)
    Cl caspase 8 (18 kDa)
    Cl caspase 9
    bactin
  • 8. IL1 is sufficient/required to inhibit TRAIL-induced apoptosis
  • 9. Vitamin D3 restores TRAIL induced apoptosis in VDR
    dependent manner
    A
    C
    Trail-THP1-VDR-/--D3
    Trail-THP1-D3-IL1
    Trail-THP1-VDR-/-
    Trail-THP1-D3
    Trail-THP1
    CTRL
    Trail
    Cl caspase 8 (41,43 kDa)
    Cl caspase 8 (18 kDa)
    B
    Cleaved caspase 9
    PARP (FL)
    PARP (Cleaved)
    Total -catenin (LE)
    Total -catenin (HE)
    actin
  • 10. CTRL
    TRAIL
    Trail/THP1
    Trail/IL1
    neo
    dnIkB
    dnAKT
    dnTCF4
    Macrophage/IL1 inhibit TRAIL-induced apoptosis through NFkB/Wnt signaling
  • 11. b catenin, an important target of TRAIL signaling
    Neo
    dnIB
    dnAKT
    dnTCF4
    Trail-THP1
    Trail–THP1
    Trail-THP1
    Trail-THP1
    Trail–IL1
    Trail-IL1
    Trail-IL1
    Trail-IL1
    CTRL
    CTRL
    CTRL
    CTRL
    Trail
    Trail
    Trail
    Trail
    Cleaved caspase 8
    Cleaved caspase-9
    -catenin
    PARP (FL)
    PARP (Cleaved)
    bactin
  • 12. Neo
    dnAKT
    dnIkB
    dnTCF4
    CTRL
    CTRL
    THP1
    THP1
    CTRL
    CTRL
    TNF
    THP1
    THP1
    TNF
    TNF
    IL1
    IL1
    TNF
    IL1
    IL1
    Snail
    c-Jun
    bactin
    Macrophages stabilize Snail in tumor cells
    THP1-STAT1 siRNA
    THP1-IL1 siRNA
    THP1-D3
    CTRL
    THP1
    THP1
    CTRL
    Snail
    Snail
    bactin
    bactin
  • 13. IL1b mediates the crosstalk between tumor cells
    and macrophages
    ??
    Snail
    Wnt targets
    b-catenin
    TCF
    STAT1
    pSTAT1
    GSK3b
    NFkB
    AKT
    IL-1b
    Proliferation
    Survival
  • 14. Versican
    Normal (71) Breast, colon, esophageal, hepatocellular, lung (235) Thyroid (35)
    Yu et al, PloS Genet 2008
    Rectal cancer Colon cancer
  • 15. VCAN stimulates macrophages to release IL1b
    A
    siRNA
    NSP
    VERSICAN
    B
    Top-Luc
  • 16. Snail regulates the expression of VCAN
    NSP Snail VER (siRNA)
    VCAN
    bcatenin
    (siRNA)
  • 17. Co-expression of VCAN and Snail in colon cancer patients
    Lin, Clinical Cancer research, 2007 (55 colon cancer patients, Affymetrics)
  • 18. IL1b: a messenger that perpetuates the crosstalk between colon cancer cells
    and macrophages
    VCAN
    EMT
    VCAN
    Vit D3
    Growth
    c-myc
    ??
    Snail
    Wnt targets
    b-catenin
    TCF
    pSTAT1
    GSK3b
    TRAIL
    NFkB
    AKT
    PDK1
    IL-1b
  • 19. Thank you:
    Pawan Kaler, PhD
    Malika Godasi, MD
    Vincent Galea
    Leonard Augenlicht, PhD