NEWS AND VIEWS

truncated, Fyn-myristoylated version of con-
© 2006 Nature Publishing Group http://www.nature.com/naturebiotechnology
Spencer, D.M. Nat. Biotechnol. 24, 1581–1590 (2006).
stitutively active Akt1 (MF-∆Akt1) with the
3.
aim of functional optimization and targeting
to intracellular membranes for efficient local- 4.
ization in lipid rafts. Delivery of the construct
5.
to dendritic cells using a replication-defective
adenovirus (Ad-MF-∆Akt1) resulted in 6.
enhanced maturation and survival of the cells 7.
both in vitro and in vivo.
(2006).
Banchereau, J. & Steinman, R.M. Nature 392, 245–
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Reis e Sousa, C. Nat. Rev. Immunol. 6, 476–483
(2006).
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(2006).
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8. Josien, R. et al. J. Exp. Med. 191, 495–502 (2000).
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der Bruggen, P. Annu. Rev. Immunol. 24, 175–208
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(2006).

Based on previous studies of dendritic-cell

longevity and vaccine efficacy8, the authors
compared their approach with ‘gold-standard’
dendritic cells in two stringent preclinical
tumor models. Ad-MF-∆Akt1 dendritic cells
massively expanded tumor antigen–specific
T cells (as much as 30% increase in numbers
of CD8+ T cells) and were clearly superior to
alternative dendritic-cell preparations for con-
trolling the growth of established melanoma
and E.G7-OVA thymomas in mice.
Interestingly, a high frequency of melanoma-
specific T cells and therapeutic efficacy did not
correlate with melanocyte destruction (viti-
ligo), even though the chosen target antigen
TRP-2 is also expressed in melanocytes. These
observations challenge the notion that effective
cancer vaccination using self-antigens neces-
sarily coincides with autoimmunity.
Finally, translating their findings to a human
context, the authors provided in vitro data
suggesting that Ad-MF-∆Akt1-transduced
dendritic cells have a more potent adjuvant
activity than mature dendritic-cell preparations
commonly used to generate tumor-specific
cytotoxic T cells.
Clearly, the work of Park et al. is not only
of considerable biological interest but also of
potential medical significance. It reinforces
the importance of dendritic-cell life span as a
parameter to be optimized for successful vac-
cination. Although assessment of the relevance
of this approach to the human situation was
limited, the in vitro data showing generation of
human tumor–specific effector T cells are prom-
ising. Only carefully controlled clinical studies
can determine the utility of Akt1-modified
dendritic cells for cancer immunotherapy.
Prophylactic vaccination represents one of
the great success stories of modern medicine
and is now being applied in oncology9. Greater
insight into various aspects of cancer vaccina-
tion (including optimization of the antigen,
adjuvant, dose and route of application10, pre-
vention of immune escape and immunosup-
pression11, as well as incorporation of lessons
learned from successful vaccines12) will be
critical for translating preclinical findings into
human therapies.
1. Nestle, F.O., Banchereau, J. & Hart, D. Nat. Med. 7,
761–765 (2001).
2. Park, D., Lapteva, N., Seethammagari, M., Slawin, K.M. &
Profile of a bacterial tumor killer
Neil S Forbes
The genome sequence of an engineered Clostridium strain will likely
facilitate the development of bacterial cancer therapies.
Common bacteria, engineered to safely seek
out and kill tumors, may well be the next gen-
eration of cancer therapeutics. In this issue,
Bettegowda et al.1 report the genome sequence
of one such bacterium, Clostridium novyi-NT,
which has been engineered to be non-toxic by
deleting the major systemic toxin gene and
has been shown to eradicate tumors in mice2.
Sequencing of this genome is a critical step
in the development of effective bacterial can-
cer therapies3. The authors also show that C.
novyi-NT spores contain mRNA that codes for
redox proteins and that vegetative C. novyi-NT
produce lipases that enable them to thrive in
the tumor environment.
Many human tumors contain regions that
are hypoxic, apoptotic and/or quiescent, and
are inaccessible to conventional cancer drugs.
Cancer cells in these regions do not prolifer-
ate and do not respond to drugs that target
rapidly growing cells4. The lack of oxygen also
limits the effectiveness of radiation therapy,
which relies on the formation of oxygen radi-
cals to damage the DNA of mitotic cells.
As cancer therapeutics, bacteria have an
advantage because they can migrate to regions
of tumors that passive drugs do not effec-
tively treat (Fig. 1). Once localized to these
regions, bacteria kill cancer cells by compet-
ing for limited nutrients and excreting cyto-
toxic agents. Bacteria can also be genetically
modified to express proteins that are toxic
to cancer cells5. In addition to Clostridium,
many other genera of bacteria have been
shown to specifically accumulate in tumors,
Neil S. Forbes is in the Department of Chemical
Engineering, The University of Massachusetts,
159 Goessmann Hall, Amherst, Massachusetts
01003, USA.
email: forbes@ecs.umass.edu
including Salmonella6–8, Bifidobacterium9
and Escherichia10.
Bacteria are not expected to replace tradi-
tional cancer drugs. Rather, the two forms of
therapy might be administered together to
more effectively kill all the cells in a tumor.
For example, complete tumor eradication
was achieved in ~60% of tumor-bearing mice
when C. novyi-NT and mitomycin C were
administered in combination2. When either C.
novyi-NT or mitomycin C was administered
alone, no tumors were destroyed. Combination
therapies should reduce the duration of cancer
treatment, lower the chance of local recurrence
and metastasis, and reduce cancer mortality.
In light of the results reported by
Bettegowda et al., I have updated a published
set of criteria for a successful bacterial cancer
therapy3. A bacterial therapeutic should (i) be
nontoxic, (ii) selectively target tumors, (iii)
target tumor regions that are unaffected by
standard therapies, (iv) not be immediately
cleared by the immune system, (v) specifically
replicate in tumors and not normal tissue,
(vi) kill cancer cells in a controllable manner,
(viii) be efficiently cleared from the body and
(ix) be genetically modifiable.
These rules paint a picture of how engi-
neered bacteria should behave in the body
(Fig. 1). First, they must have mechanisms
to seek out tumors after being injected into
the bloodstream. Next, they must localize to
regions of tumors that cannot be treated with
cancer drugs. Then, and only then, the bac-
teria must replicate to establish a substantial
population that will kill cancer cells in con-
cert with a drug present in the blood.
The work of Bettegowda et al. elucidates
some of the mechanisms that enable C. novyi-
NT to target hypoxic regions in tumors. Based
on the genomic sequence, the authors created

1484 VOLUME 24 NUMBER 12 DECEMBER 2006 NATURE BIOTECHNOLOGY

 NEWS AND VIEWS

In comparison, facultative anaerobes, such

as Salmonella typhimurium and Escherichia
coli, do not specifically target hypoxic regions

Hypoxia
and are attracted to small molecules released
Clostridium spore by quiescent and apoptotic cancer cells13,
Drug molecule which are present in many small tumors (Fig.
Active Clostridium 1). Facultative anaerobes can be administered
as active motile bacteria that can penetrate

Quiescence/apoptosis
tumor tissue in the process of localizing to
© 2006 Nature Publishing Group http://www.nature.com/naturebiotechnology

quiescent and apoptotic tumor regions. This

difference implies that the targeting mecha-
Facultative anaerobe
nisms of facultative anaerobes can be tuned,
whereas the mechanism of obligate anaerobe
targeting is fixed.
These different targeting mechanisms are
complementary, suggesting that obligate and
facultative anaerobic bacterial cancer thera-
pies should be developed in parallel. It is likely
that each bacterial genus will be better suited
Figure 1 Targeting of obligate and facultative anaerobes to tumors. After intravenous administration,
for a different tumor type and location. A fac-
both obligate and facultative anaerobes target tumors (illustrated here with a liver tumor). Obligate
anaerobes (such as Clostridium) are injected as spores and target the tumor by germinating only in low- ultative anaerobe, such as S. typhimurium,
oxygen environments (hypoxic region, blue shading). Facultative anaerobes are injected as motile active might be used to prevent metastases, whereas
bacteria, which migrate by chemotaxis and preferentially replicate in quiescent and apoptotic regions C. novyi-NT might be deployed to eradicate
of the tumor (orange shading). The figure exaggerates the distinction between hypoxic and quiescent/ large, untreatable and inoperable tumors.
apoptotic regions, which often overlap in tumors. Overall, engineered bacteria have great
promise as cancer therapeutics. By com-
pletely sequencing the C. novyi-NT genome,
gene expression arrays to determine which acid and lipid metabolism. This suggests Bettegowda et al. have taken a significant step
genes are expressed in the different phases of another mechanism that enables it to pro- towards the development of bacterial cancer
this microorganism’s life cycle. Although it liferate in necrotic tumors, which contain therapies.
was believed that bacterial spores do not con- degraded lipid membranes. 1. Bettegowda, C. et al. Nat. Biotechnol. 24, 1573–
tain RNA11, they found that spores contain One of the advantages of C. novyi-NT as 1580 (2006).
significant levels of mRNA. a cancer therapeutic is also a limitation: it is 2. Dang, L.H., Bettegowda, C., Huso, D.L., Kinzler,
K.W. & Vogelstein, B. Proc. Natl. Acad. Sci. USA 98,
A large number of genes encoded by spore an obligate anaerobe that germinates only 15155–15160 (2001).
mRNA were not expressed in growing cells. in hypoxic regions of tumors12, and cannot 3. Jain, R.K. & Forbes, N.S. Proc. Natl. Acad. Sci. USA
98, 14748–14750 (2001).
Most of these spore-specific genes encoded target nonhypoxic areas. Large tumors often 4. Brown, J.M. & Giaccia, A.J. Cancer Res. 58, 1408–
either spore coat proteins or proteins with contain hypoxic foci, but small tumors and 1416 (1998).
redox activity. The authors speculate that metastases less than 200 µm in diameter 5. Theys, J. et al. Appl. Environ. Microbiol. 65, 4295–
4300 (1999).
the redox proteins would aid germination by are often not hypoxic. The presence of oxy- 6. Zhao, M. et al. Proc. Natl. Acad. Sci. USA 102,
scavenging reactive oxygen species. Vegetative gen in small tumors and metastases would 755–760 (2005).
Clostridium cells cannot survive in environ- prevent germination of Clostridium spores. 7. Low, K.B. et al. Nat. Biotechnol. 17, 37–41
(1999).
ments that contain oxygen, which explains Another limitation of obligate anaerobes is 8. Pawelek, J.M., Low, K.B. & Bermudes, D. Cancer Res.
why C. novyi-NT is effective at targeting that they must be administered as spores, 57, 4537–4544 (1997).
tumors: it germinates and becomes active which requires that the bacteria germinate 9. Fujimori, M., Amano, J. & Taniguchi, S. Curr. Opin.
Drug Discov. Devel. 5, 200–203 (2002).
only in the strictly hypoxic regions of tumors only in tumor blood vessels that are suffi- 10. Yu, Y.A. et al. Nat. Biotechnol. 22, 313–320
and not in normal, oxygenated tissue. ciently hypoxic (Fig. 1). This limitation pro- (2004).
The gene expression analysis of Bettegowda hibits obligate anaerobes from ‘searching’ 11. Liu, H. et al. J. Bacteriol. 186, 164–178 (2004).
12. Lambin, P. et al. Anaerobe 4, 183–188 (1998).
et al. also revealed that when C. novyi-NT for hypoxic regions by penetrating through 13. Kasinskas, R.W. & Forbes, N.S. Biotechnol. Bioeng.
infects tumors, it expresses genes for fatty interstitial tumor tissue. 94, 710–721 (2006).

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