The Farnesyltransferase Inhibitors Tipifarnib and Lonafarnib
Inhibit Cytokines Secretion in a Cellular Model of Mevalonate
Kinase Deficiency
ANNALISA MARCUZZI, LUIGINA DE LEO, GIULIANA DECORTI, SERGIO CROVELLA, ALBERTO TOMMASINI,
AND ALESSANDRA PONTILLO
Medical Genetic Service [A.M., S.C., A.P.], Laboratory of Immunopathology [A.T.], Institute for Maternal and Child Health “Burlo
Garofolo,” Trieste 34134, Italy; Department of Reproductive and Developmental Sciences and Public Health Care [L.D.L.], Department
of Life Sciences [G.D.], University of Trieste, Trieste 34137, Italy; Department of Genetics [S.C.], Federal University of Pernambuco,
Cidade Universitaria, 50.740-530 Recife, Brazil
ABSTRACT: The shortage of geranylgeranyl-pyrophosphate
(GGPP) was associated to an increased IL-1
release in the autoin-
flammatory syndrome mevalonate kinase deficiency (MKD), a rare
inherited disease that has no specific therapy. Farnesyltransferase
inhibitors (FTIs) act at the end of mevalonate pathway. Two FTIs,
Tipifarnib (Tip) and Lonafarnib (Lon), were therefore evaluated as
possible therapeutical choices for the treatment of MKD. FTIs could
lead to a redirection of the limited available number of mevalonate
intermediates preferentially to GGPP synthesis, eventually prevent￾ing the uncontrolled inflammatory response. The effect of Tip and
Lon on intracellular cholesterol level (ICL) and on proinflammatory
cytokines secretion was evaluated in a cellular model of MKD,
chemically obtained treating RAW 264.7 cells with lovastatin (Lova)
and alendronate (Ald). The combination of FTIs with the isoprenoid
geraniol (GOH) was also tested both in this model and in monocytes
isolated from MKD patients. Tip and Lon proved to revert the ICL
lowering and to significantly reduce the lipopolysaccharide-induced
cytokines secretion in Ald-Lova -RAW 264.7 cells. This anti￾inflammatory effect was amplified combining the use of GOH with
FTIs. The effect of GOH and Tip was successfully replicated in MKD
patients’ monocytes. Tip and Lon showed a dramatic anti￾inflammatory effect in monocytes where mevalonate pathway was
chemically or genetically impaired. (Pediatr Res 70: 78–82, 2011)
n the familial mevalonate kinase deficiency (MKD, OMIM:
*251170), the second enzyme of the mevalonate pathway
[mevalonate kinase, MK/mevalonate kinase gene
(NM_000431) (MVK)] is mutated and shows a reduced activ￾ity, leading to a shortage of downstream compounds. In
particular, the lack of geranylgeranyl-pyrophosphate (GGPP)
results in an augmented caspase-1-dependent IL-1
secretion
that is the major cytokine responsible for the inflammatory
systemic effects observed in MKD patients (1,2). These sub￾jects present periodic fever attacks associated with lymphade￾nopathy, abdominal, articular, and cutaneous involvement,
and, in most severe cases, also neurological impairment (3).
Despite the many efforts done in the past decades to elucidate
the molecular events linking the mevalonate pathway impair￾ment to the inflammatory clinical phenotype, no specific
treatment has been yet developed for MKD.
We and other authors have recently proposed natural exog￾enous isoprenoids (NEIs) as a possible therapeutic approach
for MKD. These compounds, because of their isoprenoid
structure, are supposed to enter the mevalonate pathway and to
bypass the biochemical block, reconstituting the pathway and
limiting the shortage of GGPP (2,4,5) (Fig. 1).
With similar results, the farnesyltransferase inhibitor (FTI)
Manumycin A (ManA) was recently used in our cellular and
murine model of MKD (6). FTIs are a class of experimental
drugs that target protein FT with the downstream effect of
preventing the proper functioning of Ras protein, and they
have been studied as anticancer agents but have not been
definitely approved yet (7,8). In our opinion, FT inhibition in
MKD cells lead to a redirection of the limited available
number of GGPP molecules preferentially to geranylgerany￾lation, thereby preventing the activation of caspase-1 and
production of mature IL1
(Fig. 1).
Taking into account all these findings, and in particular
those obtained with ManA, we decided to investigate the
effect of two novel FTIs, Tipifarnib (Tip) and Lonafarnib
(Lon) (9,10), on the inflammation induced by the inhibition of
the mevalonate pathway, in the hypothesis to find out an
alternative use for these anticancer drugs in the treatment of
MKD (Fig. 1).
Tip and Lon were first tested in a cellular model of MKD
obtained treating the murine monocytic cell lines RAW 264.7
with two known inhibitors of the mevalonate pathway, alen￾dronate (Ald) (11) and lovastatin (Lova) (12), and then in
monocytes isolated from two MKD patients. The cellular
model obtained with the combined use of Ald and Lova aims
Received October 7, 2010; accepted January 12, 2011.
Correspondence: Annalisa Marcuzzi, Ph.D., Medical Genetic Service, Institute for
Maternal and Child Health “Burlo Garofolo,” Via dell’Istria 65/1, 34134 Trieste, Italy;
e-mail: [email protected]
Supported by a grant from the Institute for Maternal and Child Health IRCCS “Burlo
Garofolo,” Italy (RC 13/2008).
Abbreviations: Ald, alendronate; FTIs, farnesyltransferase inhibitors;
GGPP, geranylgeranyl-pyrophosphate; GOH, geraniol; ICL, intracellular
cholesterol level; Lon, Lonafarnib, Sarasar; Lova, lovastatin; LPS, lipopoly￾saccharide; ManA, Manumycin A; MK, mevalonate kinase (E.C. 2.7.1.36);
MKD, mevalonate kinase deficiency; MTT, 3-(4.5-dimethylthiazol-2-yl)-2.5-
diphenyltetrazolium bromide; MVK, mevalonate kinase gene (NM_000431);
NEIs, natural exogenous isoprenoids; Tip, Tipifarnib, Zarnestra
0031-3998/11/7001-0078
PEDIATRIC RESEARCH
Vol. 70, No. 1, 2011
Copyright © 2011 International Pediatric Research Foundation, Inc.
Printed in U.S.A.
78
to reproduce as faithfully as possible the condition of block
metabolic pathway of cholesterol. The results were compared
with those obtained with NEIs for their anti-inflammatory
action that has been recently described in these models (6,13):
finally, we evaluated the additive effect of FTIs and NEIs.
MATERIALS AND METHODS
Chemicals. Lipopolysaccharide (LPS, E. coli-serotype 055:B5, 1 mg/mL
stock in H2O), Ald (30 mM), geraniol (GOH, 6 M; Euphar group s.r.l.,
Piacenza, Italy), and Lova (50 mM) were dissolved in saline solution. ManA
(10 M), Tip (Tip, 20 mM), and Lon (16.5 mM) were dissolved in DMSO so
that the final concentration of DMSO would not exceed 0.1%. Tip (Tip,
R115777, Zarnestra) and Lon (SCH66336, Sarasar) were kindly provided by
Prof G. Martinelli (Institute of Hematology ‘L and A Sera`gnoli’, University
of Bologna, Bologna, Italy). All the reagents were from Sigma Chemical
Co.-Aldrich (Milan, Italy), except where differently specified.
RAW 264.7 cell culture. RAW 264.7 cells (murine monocyte/macrophage
cell line) were cultured in DMEM supplemented with 10% fetal bovine serum
with 100 M Ald and 20 M Lova for 20 h and then with 10 g/mL LPS for
additional 24 h. At the end of the incubation period, the supernatants were
collected for cytokine evaluation, while the cells were pelleted for the
quantification of intracellular cholesterol. In the experiments, GOH (100 M),
ManA (10 M), Tip (5 M), or Lon (5 M) were added together with Ald
and Lova.
MKD patients and monocyte isolation. Monocytes isolated from the
peripheral blood of two MKD subjects (patient 1, P1: 4 y old, male, MVK:
S135L/V377I; patient 2, P2: 10 y old, male, MVK: I268T/V377I), through
standard protocol, were cultured at a cell density of 2
105 cells/well in
RPMI 1640 containing 10% fetal bovine serum (Euroclone, Milan, Italy).
Cells were incubated with 10 M GOH for 20 h, or 5 M Tip for 1 h, or their
combination. Cells were then treated with 1 g/mL LPS for supplementary
24 h. At the end of the incubation period, the supernatants were collected for
IL1
assay. Written informed consent was obtained from patients’ parents,
according to the protocol of the ethical board of Institute of Children Health
“Burlo Garofolo” (Trieste, Italy; n.185/08, 19/08/2008).
Determination of cytokines release (IL1
, IL18, and TNF). IL1
, IL18,
and TNF concentrations were determined in the cell culture medium (su￾pernatant) by ELISA kits according to manufacturer’s protocols, and the
amount of cytokines expressed as picograms per milliliters (Endogen Human
IL-1
ELISA Kit; Pierce Biotechnology Inc., Rockford, IL).
Determination of intracellular free cholesterol. The amount of free
cholesterol was determined with the Amplex red cholesterol assay kit accord￾ing to the manufacturer’s instructions (Molecular Probes, Invitrogen), and the
amount of cholesterol expressed as micrometers (14,15).
Evaluation of cytotoxicity. Tip, Lon, ManA, and GOH toxicity were
evaluated with the MTT (3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyltetrazo￾lium bromide) assay according to the method of Mosmann (16). Briefly, 2

105
/mL RAW 264.7 cells or human monocytes were incubated with 10 to 200
M Ald, 5 to 40 M Lova, 1 to 50 g/mL LPS, 1 to 20 M Tip, 1 to 20 M
Lon, 1 to 20 M ManA, and GOH 10 to 100 M alone and in combination,
for 3 d; before adding MTT reagent.
Statistical analysis. All results are expressed as the mean SE. Statistical
significance was calculated using one-way ANOVA, followed by Tukey
multiple comparison test. p values less than 0.05 were considered significant.
Statistical analysis was performed using the GraphPad Prism software version
5 (GraphPad Software, San Diego, CA).
RESULTS
Tip (1–20 M) or Lon (1–20 M) (9,17,18) were added to
the cellular model of MKD obtained treating RAW 264.7 cells
simultaneously with 100 M Ald and 20 M Lova. Ald
Lova lead to a significantly reduction of intracellular choles￾terol level (ICL; 10.43 1.12 M) when compared with
untreated cells (14.40 0.72 M) as expected being inhibi￾tors of mevalonate pathway; this inhibition was amplified in
the presence of LPS (Ald Lova LPS: 7.88 0.28 M).
In Figure 2, the effect of 5 M Tip and 5 M Lon was
reported as minor dose with maximum results. We tested
several concentration of both compounds and finally we chose
the dose of 5 M according to indication in literature data.
The DMSO used as solvent for the two compounds did not
alter any of the results (data not showed).
Tip and Lon were able to significantly revert the diminish￾ing of ICL (9.70 0.17 M and 9.76 0.40 M, respec￾tively) compared with Ald Lova LPS. These results were
comparable with those obtained with ManA (9.53 0.56 M)
and GOH (9.6 0.69 M).
The secretion in the cell culture medium of proinflamma￾tory cytokines IL1
, IL18, and TNF were significantly re￾duced in Ald-Lova-LPS cells treated with Tip and Lon (Table
1). In Figure 3, the effect of Tip (5 M) and Lon (5 M) on
IL1
reduction was reported as exemplificative of the three
cytokines. Similar reductions (Tip: 44.3%; Lon: 46%) were
obtained with the natural isoprenoid GOH (54.3%) or with
another FTI, ManA (32.3%; Fig. 3). The cytokine release to
Figure 1. Schematic representation of mevalonate pathway. Compounds
used in the experiments are indicated along the pathway in bold characters.
Figure 2. Tipifarnib and Lonafarnib rescue the levels of intracellular cho￾lesterol in Ald-Lova-LPS RAW 264.7 cells. Cells were incubated with 100
M alendronate (Ald) and 20 M lovastatin (Lova) for 20 h, and then with
10 g/mL LPS for supplementary 24 h. Bars represent the means ICL
(M) SE of three independent experiments. *p  0.05 significantly
different from ICL in Ald Lova LPS-treated cells. **p  0.01 signifi-
cantly different from ICL in Ald Lova LPS-treated cells.
FARNESYLTRANSFERASE INHIBITORS IN MKD 79
the untreated cells (230 20 pg/mL) and in presence of Ald
and Lova (316 20 pg/mL) alone can be seen in the Figure
3. Tip and Lon were able to rescue the IL-1
secretion
compared with LPS-treated cells even if not in a statistically
significant way (Fig. 3). When GOH was used together with
Tip and Lon, the effect of the two FTIs on the IL1
, IL18, and
TNF secretion was amplified (Table 2) as reported in Figure 4
for IL1
.
Tip GOH and Lon GOH significantly diminish IL1

secretion (277.1 92.1 pg/mL and 421.9 122.5 pg/mL,
respectively) when compared with Ald Lova LPS-treated
cells (5519 1497 pg/mL; p  0.05) as well as to Tip
(2291 453.2 pg/mL; p  0.05), Lon (2305 359.8 pg/mL;
p  0.05), or GOH (2811 271.1 pg/mL; p  0.05) alone
(Fig. 4). A similar additive anti-inflammatory effect was also
observed using GOH combined with ManA (Fig. 4).
The combination Tip GOH resulted significantly more
effective than Lon GOH (p  0.001), and so we decided to
test Tip, GOH, and Tip GOH on LPS-induced IL-1

secretion in monocytes isolated from two MKD patients. In
MKD patient 1 (P1), Tip was more effective in diminishing
IL-1
secretion compared with LPS-treated cells alone or in
combination with 10 M GOH even if not in a statistically
significant way (Tip LPS 107.8 32.8 pg/mL; Tip
GOH LPS 92.36 0.47 pg/mL versus LPS 171.9 11.6
pg/mL: p; Fig. 5A). This effect was registered also in MKD
patient 2 (P2): Tip and Tip GOH significantly reduced
LPS-induced IL-1
secretion in (P2) (Tip LPS 273.3 9.2
pg/mL, Tip GOH LPS 188.2 20.5 pg/mL versus LPS
1068 92.7 pg/mL: p  0.001; Fig. 5B). Tip and Lon, at the
concentration used, did not result cytotoxic when evaluated
with the MTT assay (data not shown).
DISCUSSION
The inhibition of farnesyltransferase is nowadays a com￾mon target for antineoplastic drugs because of the consequent
reduction of farnesylated protein such as small GTP proteins
of oncogene Ras family (19). Recently, we reported the use of
the FTI ManA as a possible choice for the treatment of the
hereditary rare and still orphan disease MKD (6). The hypoth￾esized mechanism of FTIs is principally related to the redi￾rection of GGPP to geranylgeranylation (6).
In this study, we decided to investigate the effect of other
two FTIs, the Tip and Lon, that are currently used in clinical
trial (20,21) in a MKD cellular model because in our opinion,
they could be more easily and rapidly included in the treat￾ment of other inflammatory diseases.
In this cellular model, the combination of Ald and Lova
induced an expected and significant reduction in intracellular
cholesterol, similarly to what shown in mice serum level (22),
and let us using this parameter to verify the changes in
mevaloante pathway induced by different compounds (iso￾prenoids or FTIs).
Tip and Lon were able to revert the decrease of intracellular
cholesterol induced by Lova and Ald (Fig. 2; inhibitors of
HMG-CoA-red and farnesyl-pyrophosphate synthase, respec￾tively, as shown in Fig. 1), demonstrating that these com￾pounds, acting on FT, could modulate the outcome of the
residual intermediates of mevalonate pathway below farne￾sylpyrophosphate synthase driving them through the other two
main exits, cholesterol biosynthesis and geranylgeranylation.
Moreover, these FTIs reduced proinflammatory cytokines
secretions either in MKD cellular model (Fig. 3) and in
monocytes isolated from MKD patients (Fig. 5), emphasizing
the hypothesis that the reequilibration of GGPP production
could ameliorate the inflammatory state because of a biochem￾ical or a familial unpaired mevalonate pathway (6). Similar
results were previously obtained through the use of exogenous
isoprenoids, such as GOH, farnesol, and geranylGOH (4,13),
Table 1. Effect of Tipifarnib (Tip), Lonafarnib (Lon), Manumycin A (ManA), and Geraniol (GOH) on LPS-induced cytokines secretion in
2329 195.2 0.01* 2329 195.2 0.01*
Data were representative of three distinct experiments. Mean concentration and SE are reported.
* Significantly different from cytokine concentration in Ald-Lova-LPS cells.
† Significantly different from cytokine concentration in Ald-Lova-LPS cells.
Figure 3. Tipifarnib and Lonafarnib are able to lower the IL-1
secretion in
Ald-Lova-LPS RAW 264.7 cells. Cells were incubated with 100 M alen￾dronate (Ald) and 20 M lovastatin (Lova) for 20 h, and then with 10 g/mL
LPS for supplementary 24 h (Ald Lova LPS). FTIs (10 M ManA, 5 M
Tip, 5 M Lon) and GOH (100 M) were added simultaneously with Ald and
Lova. Bars represent the mean concentration (pg/mL) SE of three exper￾iments. *p  0.05 significantly different from cytokine concentration in
Ald-Lova-LPS cells. **p  0.01 significantly different from cytokine con￾centration in Ald-Lova-LPS cells.
80 MARCUZZI ET AL.
and they were replicated with GOH in this study to compare
the two different anti-inflammatory approaches.
It seems that the anti-inflammatory effect of Tip and Lon, as
well as of GOH, was independent from the type of secreted
cytokine, IL-1
, IL-18, or TNF, indicating a mechanism
acting before the specific and different inflammatory signaling
pathways. IL-1
and IL-18 were produced after the caspase-1
activation, an event specifically induced by the inhibition of
mevalonate pathway as reported by Mandey et al (2). For what
concern the inhibition on TNF secretion, no theory has been
yet proposed, even if previous works showed that the block of
mevalonate pathway induced TNF secretion from monocytes
underling the causative link between cholesterol metabolism
and TNF (23). We hypothesize that a small GTP protein or a
prenilated protein could be involved in the signaling leading to
TNF production, similarly to what happens with the activation
of caspase-1 (2).
We first tried, unsuccessfully, to decrease the cytokines
secretions to the basal condition with augmenting concentra￾tion of both FTIs and GOH, considering their similar anti￾inflammatory effect (data not shown). We then tried with the
combination of Tip or Lon and GOH: this significantly re￾duced the cytokines production (Fig. 4). Beyond expectation,
FTIs and GOH showed a synergic effect, probably related to
the bypassing of a compensative dose-related mechanism
induced by each single compound, or alternative to the dif￾ferent mechanism of action of FTIs and GOH.
Our results suggest that the manipulation of the mevalonate
pathway through the use of FTIs or GOH with the purpose to
rebalance the intermediates levels acts above and in an aspe￾Figure 4. GOH enhances the anti-inflammatory effect of FTIs. Cells were
incubated with 100 M alendronate (Ald) and 20 M lovastatin (Lova)
(Ald Lova) for 20 h, and then with 10 g/mL LPS for supplementary 24 h.
FTIs (10 M ManA, 5 M Tip, 5 M Lon) and 100 M GOH were added
simultaneously with Ald and Lova. Bars represent the mean concentration
(pg/mL) SE of three experiments. **p  0.01 significantly different from
cytokine concentration in Ald-Lova-LPS cells.
Figure 5. Tipifarnib, alone or in combination with GOH, is able to lower the
LPS-induced IL-1
secretion in MKD monocytes. LPS-induced IL-1
secre￾tion in monocytes from two MKD patients (P1-4A; P2-4B) incubated in the
absence or presence of 10 M GOH for 20 h, or 5 M Tip for 1 h, or their
combination. Cells were then treated 1 g/mL LPS for supplementary 20 h.
Absolute values (pg/mL) were obtained from one experiment performed in
triplicate. *p  0.05 significantly different from absolute value of cytokine
concentration in LPS cells (100%).
IL-1
(pg/mL) 2811 157.1 2673 305.8 264.5 3.94 p  0.05 p  0.05
IL-18 (pg/mL) 730.6 17.77 611.6 6.3 118.9 13.88 p  0.01 p  0.05
TNF- (pg/mL) 1841 44.84 2329 195.2 394.1 81.49 p  0.01 p  0.01
Data were representative of three distinct experiments. Mean concentration and SE are reported.
P1, p value of the comparison between Ald Lova LPS GOH Tip respect and ALD Lova LPS GOH; P2, p value of the comparison between
Ald Lova LPS GOH Tip respect and ALD Lova LPS Tip; P3, p value of the comparison between Ald Lova LPS GOH Lon respect
and ALD Lova LPS GOH; P4, p value of the comparison between Ald Lova LPS GOH Lon respect and ALD Lova LPS Lon; P5,
p value of the comparison between Ald Lova LPS GOH ManA respect and ALD Lova LPs GOH; P6, p value of the comparison between
Ald Lova LPS GOH ManA respect and ALD Lova LPS ManA.
FARNESYLTRANSFERASE INHIBITORS IN MKD 81
cific mechanism. Tip used in monocytes isolated from two
MKD patients showed interesting results (Fig. 5) even if a
certain interindividual variability and the limited number of
patients did not allow us to definitively demonstrate the
efficacy ex vivo of this compound.
The different entity of response between the two patients
could be explained taking in account the interaction between
genetic defect and compounds concentration. In MKD, resid￾ual MK activity varies from 0.5 to 7% depending on the type
of MVK mutations (24). Considering that MK activity affects
the levels of GGPP and the consequent shortage of gera￾nylgeranylation, monocytes from subjects carrying different
MVK mutations could need different amount of Tip to exhibit
the same anti-inflammatory effect. A quantitative residual MK
activity evaluation was not performed in our patients because
it is quite expensive and not so informative for the clinical
follow-up, but the possible residual MK activity could at least
in part explain the different response to isoprenoids in the two
patients carrying different mutations.
In conclusion, in agreement with previously reported data
(2,4,6), our findings suggest once more that the recovery of
GGPP flux through the geranylgeranyltransferase, in this case
through the use of Tip and Lon, has an anti-inflammatory
effect, both in pharmacologic or in genetic blockage of the
mevalonate pathway. These molecules are nowadays used in
clinical protocols as anticancer drugs, so when considering our
findings and those already reported in the literature, we would
like to propose the use of Tip and Lon as a novel therapeutical
approach for the still orphan disease MKD.
Acknowledgments. We thank Prof G. Martinelli and Dr. I.
Iacobucci (Institute of Hematology ‘L and A Sera`gnoli’,
University of Bologna, Bologna, Italy) for giving us Tipi￾farnib and Lonafarnib.
REFERENCES
1. Kuijk LM, Mandey SH, Schellens I, Waterham HR, Rijkers GT, Coffer PJ, Frenkel J
2008 Statin synergizes with LPS to induce IL-1beta release by THP-1 cells through
activation of caspase-1. Mol Immunol 45:2158 –2165
2. Mandey SH, Kuijk LM, Frenkel J, Waterham HR 2006 A role for geranylgerany￾lation in interleukin-1beta secretion. Arthritis Rheum 54:3690 –3695
3. Haas D, Hoffmann GF 2006 Mevalonate kinase deficiencies: from mevalonic
aciduria to hyperimmunoglobulinemia D syndrome. Orphanet J Rare Dis 1:13
4. Marcuzzi A, Pontillo A, De Leo L, Tommasini A, Decorti G, Not T, Ventura A 2008
Natural isoprenoids are able to reduce inflammation in a mouse model of mevalonate
kinase deficiency. Pediatr Res 64:177–182
5. Frenkel J, Rijkers GT, Mandey SH, Buurman SW, Houten SM, Wanders RJ,
Waterham HR, Kuis W 2002 Lack of isoprenoid products raises ex vivo interleukin-
1beta secretion in hyperimmunoglobulinemia D and periodic fever syndrome.
Arthritis Rheum 46:2794 –2803
6. De Leo L, Marcuzzi A, Decorti G, Tommasini A, Crovella S, Pontillo A 2010
Targeting farnesyl-transferase as a novel therapeutic strategy for mevalonate kinase
deficiency: in vitro and in vivo approaches. Pharmacol Res 61:506 –510
7. Appels NM, Beijnen JH, Schellens JH 2005 Development of farnesyl transferase
inhibitors: a review. Oncologist 10:565–578
8. Sebti SM 2005 Protein farnesylation: implications for normal physiology, malignant
transformation, and cancer therapy. Cancer Cell 7:297–300
9. Xue X, Lai KT, Huang JF, Gu Y, Karlsson L, Fourie A 2006 Anti-inflammatory
activity in vitro and in vivo of the protein farnesyltransferase inhibitor tipifarnib.
J Pharmacol Exp Ther 317:53– 60
10. Basso AD, Mirza A, Liu G, Long BJ, Bishop WR, Kirschmeier P 2005 The farnesyl
transferase inhibitor (FTI) SCH66336 (lonafarnib) inhibits Rheb farnesylation and
mTOR signaling. Role in FTI enhancement of taxane and tamoxifen anti-tumor
activity. J Biol Chem 280:31101–31108
11. Russell RG, Watts NB, Ebetino FH, Rogers MJ 2008 Mechanisms of action of
bisphosphonates: similarities and differences and their potential influence on clinical
efficacy. Osteoporos Int 19:733–759
12. Hilgendorff A, Muth H, Parviz B, Staubitz A, Haberbosch W, Tillmanns H,
Holschermann H 2003 Statins differ in their ability to block NF-kappa B activation
in human blood monocytes. Int J Clin Pharmacol Ther 41:397– 401
13. Marcuzzi A, Tommasini A, Crovella S, Pontillo A 2010 Natural isoprenoids inhibit
LPS-induced-production of cytokines and nitric oxide in aminobisphosphonate￾treated monocytes. Int Immunopharmacol 10:639 – 642
14. Frey T, De Maio A 2007 Increased expression of CD14 in macrophages after
inhibition of the cholesterol biosynthetic pathway by lovastatin. Mol Med 13:592–
604
15. Iftakhar-E-Khuda I, Koide N, Hassan F, Noman AS, Dagvadorj J, Tumurkhuu G,
Naiki Y, Komatsu T, Yoshida T, Yokochi T 2009 Novel mechanism of U18666A￾induced tumour necrosis factor-alpha production in RAW 264.7 macrophage cells.
Clin Exp Immunol 155:552–558
16. Mosmann T 1983 Rapid colorimetric assay for cellular growth and survival:
application to proliferation and cytotoxicity assays. J Immunol Methods 65:55– 63
17. Desrosiers RR, Cusson MH, Turcotte S, Beliveau R 2005 Farnesyltransferase
inhibitor SCH-66336 downregulates secretion of matrix proteinases and inhibits
carcinoma cell migration. Int J Cancer 114:702–712
18. O’Meara SJ, Kinsella BT 2005 The effect of the farnesyl protein transferase inhibitor
SCH66336 on isoprenylation and signalling by the prostacyclin receptor. Biochem J
386:177–189
19. Braun T, Fenaux P 2008 Farnesyltransferase inhibitors and their potential role in
therapy for myelodysplastic syndromes and acute myeloid leukaemia. Br J Haematol
141:576 –586
20. Venkatasubbarao K, Choudary A, Freeman JW 2005 Farnesyl transferase inhibitor
(R115777)-induced inhibition of STAT3(Tyr705) phosphorylation in human pan￾creatic cancer cell lines require extracellular signal-regulated kinases. Cancer Res
65:2861–2871
21. Baulch-Brown C, Molloy TJ, Yeh SL, Ma D, Spencer A 2007 Inhibitors of the
mevalonate pathway as potential therapeutic agents in multiple myeloma. Leuk Res
31:341–352
22. Marcuzzi A, Decorti G, Pontillo A, Ventura A, Tommasini A 2010 Decreased
cholesterol levels reflect a consumption of anti-inflammatory isoprenoids associated
with an impaired control of inflammation in a mouse model of mevalonate kinase
deficiency. Inflamm Res 59:335–338
23. Deng X, Yu Z, Funayama H, Yamaguchi K, Sasano T, Sugawara S, Endo Y 2007
Histidine decarboxylase-stimulating and inflammatory effects of alendronate in
mice: involvement of mevalonate pathway, TNFalpha, macrophages, and T-cells. Int
Immunopharmacol 7:152–161
24. Houten SM, Wanders RJ, Waterham HR 2000 Biochemical and genetic aspects of
mevalonate kinase and its deficiency. Biochim Biophys Acta 1529:19 –32
82 MARCUZZI ET AL.