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