Robert Ramer et al, 2012
University of Rostock, Germany
Presented by: Steven Addison
Cannabidiol inhibits lung cancer cell invasion and metastasis via ICAM-1
Cannabidiol (CBD)
Major alkaloid found in cannabis
Used to treat epilepsy, spasms, anxiety, schizophrenia, nausea
Found to inhibit cancer growth (Mechoulam 2007)
Found to inhibit cancer cell invasiveness (McAllister 2007)
Tetrahydrocannabinol (THC) Methanandamide (MA)
ProliferationAngiogenesis
ApoptosisAutophagy
CBD Background
Decreases ID-1: Inhibitor of differentiation via H-L-H Transcription Factors (McAllister 2007)
TIMP-1: Tissue inhibitor of matrix metalloproteinases, anti-invasive (Ramer 2010)
Signaling by CB1, CB2, TRPV1 (transient receptor potential vanilloid)
CBD, THC & MA (Ramer 2008)
ICAM-1: Intercellular adhesion molecule Upregulated by CBD in lung cancer cells
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ICAM-1 aka CD54
Immunoglobulin superfamily
80-110 kDa transmembrane glycoprotein
Expressed on cell surfaces, binds β2 integrin
Trafficking inflammatory cells, immunosurveillance
Cell-cell interactions, antigen presentation
Signal transduction – MAPK (mitogen activated protein kinase)
Lymphocyte adhesion to tumor cells
Decreased metastasis of several cancer types
May upregulate TIMP-1
Klaus Ley 2001Nature Medicine 7, 1105 - 1106
Outline of Current Study
CBD and ICAM-1, TIMP-1 in cancer cells
Receptor involvement in CBD activity – CB1, CB2, TRPV1
ICAM-1 neutralizing antibody and CBD effects
ICAM-1 siRNA knockdown and CBD effects
Other cannabinoids – THC and MA
Primary metastasized human lung tumor cells
Metastasis study in vivo – athymic mice xenograft
CBD and ICAM-1Figure 1
Concentration-dependent effect of CBD on ICAM-1 mRNA and protein expression in A549 (A), H358 (B), or H460 (C) human cancer cell lines
Time course of ICAM-1 and TIMP-1 mRNA expression of CBD- or vehicle-treated A549 (D), H358 (E), and H460 (F) cells assessed by real-time RT-PCR
CBD and ICAM-1/TIMP-1Figure 1
AM-251: CB1 antagonist
AM-630: CB2 antagonist
Capsazepine: TRPV1 antagonist
Effect of AM-251, AM-630, and capsazepine on CBD-induced ICAM-1 protein expression and p42/44 MAPK activation.
CBD signal transductionFigure 2
SB203580: inhibitor of p38 MAPK activation
PD98059: inhibitor of p42/44 MAPK activation
Effect of SB203580 and PD98059 on CBD-induced ICAM-1 protein expression.
CBD and MAPKsFigure 2
Effect of a neutralizing ICAM-1 antibody on anti-invasive and TIMP-1-inducing action of CBD
CBD’s ICAM-1 dependence
Figure 3
Effect of ICAM-1 siRNA on anti-invasive and TIMP-1-inducing action of CBD
CBD’s ICAM-1 dependence
Figure 4
Effect of THC and MA on ICAM-1 mRNA expression.
Effect of an ICAM-1 antibody on TIMP-1 expression by THC and MA.
Other cannabinoidsFigure 5
Influence of ICAM-1 siRNA on the TIMP-1-inducing action of THC and MA
Other cannabinoidsFigure 6
Effect of the cannabinoids CBD, THC, and MA on the invasiveness of primary tumor cells
Cells from patientsTable 2
Regulation of ICAM-1, TIMP-1, and invasiveness by cannabinoids in primary lung cancer cells
Cells from patientsFigure 7
CBD in vivoRole of ICAM-1 in CBD’s antimetastatic effect in athymic nude mice xenografted with A549 cells and treated with CBD every 72 hours for 28 days
Impact
Cannabinoid role in anti-invasive TIMP-1 pathway elucidated - receptor/MAPK mediated ICAM-1 upregulation discovered
ICAM-1 shown to be a key player in anti-invasive mechanism
Anticarcinogenic properties of cannabinoids demonstrated in metastasized primary tumor cells from humans
In vivo demonstration of an inhibitor-based antimetastatic mechanism elicited by cannabinoids
Future Work
CLINICAL TRIALS
Q’s and A’s