Cell Stem Cell

Previews

1995). In fact, gene expression pro-
grams activated by cell-surface receptors
impinging on Rho proteins rely on the
nuclear integration of mechanisms sensing
cytoskeletal changes with activation of
MAPK cascades (Bar-Sagi and Hall, 2000;
Posern and Treisman, 2006; Olson and
Nordheim, 2010). Thus, we can speculate
that if epithelial stem cells attach to the
ECM but are restricted from subsequent
spreading by constraints imposed by the
surface geometry, only partial deployment
of the integrin-initiated signaling network
may result, and hence lead to the uncoordi-
nated activation of Rho GTPases, MAPKs,
and cytoskeletal components. This combi-
nation of signals may result in the initiation
of cell differentiation programs, causing
cells to exit the cell cycle permanently and
thus the stem cell pool.
Although the relevance of these mecha-
nisms to in vivo stem cell maintenance
remains to be elucidated, it is known that
signaling events that mediate the interac-
tion between the basement membrane
2010). In addition, the mutant cells differ-
entiate and thus exit from the stem cell
pool, resulting in defective wound closure
(Castilho et al., 2010). We can hypothesize
that the requirement for the integration of
proliferative cues with mechanosensing
pathways in order to communicate the
condition of the niche may serve as a fail-
safe mechanism that triggers differenti-
ating gene programs when stem cells
sense aberrant biophysical conditions
(Figure 1). The depletion of epithelial stem
cells by promoting their differentiation in
response to extrinsic or intrinsic cellular
stress pathways may also act as a pro-
tective mechanism against oncogenic
perturbation of this particularly vulnerable
self-renewing cell population (Castilho
et al., 2009). Alternatively, this emerging
tendency of stem cells to undergo differen-
tiation under stress conditions may also
contribute to pathological events, such as
accelerated tissue aging or reduced tissue
regeneration. On the other hand, failure to
activate differentiation pathways may
multiple diseases characterized by defec-
tive stem cell differentiation, including
cancer, or by accelerated stem cell deple-
tion, which leads to loss of tissue regener-
ative capacity and premature aging.
REFERENCES
Bar-Sagi, D., and Hall, A. (2000). Cell 103, 227–238.
Castilho, R.M., Squarize, C.H., Chodosh, L.A., Wil-
liams, B.O., and Gutkind, J.S. (2009). Cell Stem
Cell 5, 279–289.
Castilho, R.M., Squarize, C.H., Leelahavanichkul,
K., Zheng, Y., Bugge, T., and Gutkind, J.S.
(2010). PLoS ONE 5, e10503.
Connelly, J.T., Gautrot, J.E., Trappmann, B., Tan,
D.W., Donati, G., Huck, W.T., and Watt, F.M.
(2010). Nat. Cell Biol. 12, 711–718.
Coso, O.A., Chiariello, M., Yu, J.C., Teramoto, H.,
Crespo, P., Xu, N., Miki, T., and Gutkind, J.S.
(1995). Cell 81, 1137–1146.
Ezhkova, E., Pasolli, H.A., Parker, J.S., Stokes, N.,
Su, I.H., Hannon, G., Tarakhovsky, A., and Fuchs,
E. (2009). Cell 136, 1122–1135.
Geiger, B., Spatz, J.P., and Bershadsky, A.D.
(2009). Nat. Rev. Mol. Cell Biol. 10, 21–33.

and the basal epidermal stem cells are
required to maintain their stem-like state.
For example, stem cells lacking the small
GTPase Rac1 fail to undergo shape
changes and migrate on the provisional
ECM at the wound edge (Castilho et al.,
contribute to psoriasis and other skin hy-
perproliferative conditions. Hence, the
molecular dissection of the underlying
mechanisms that exist in the epidermal
stem cell niche may provide new molecular
targets for pharmacological intervention in
Olson, E.N., and Nordheim, A. (2010). Nat. Rev.
Mol. Cell Biol. 11, 353–365.
Posern, G., and Treisman, R. (2006). Trends Cell
Biol. 16, 588–596.
Samuel, M.S., and Olson, M.F. (2010). Cell Stem
Cell 7, this issue, 135–136.

Differential DNA Damage Response

in Stem and Progenitor Cells
Jun Seita,1,* Derrick J. Rossi,2,3,4,5 and Irving L. Weissman1
1StanfordInstitute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA 94305, USA
2Harvard Stem Cell Institute, Boston, MA 02115, USA
3Department of Pathology, Harvard Medical School, Boston, MA 02115, USA
4Immune Disease Institute, Boston, MA 02115, USA
5Program in Cellular and Molecular Medicine, Children’s Hospital Boston, Boston, MA 02115, USA

*Correspondence: jseita@stanford.edu
DOI 10.1016/j.stem.2010.07.006

The long lifespan of tissue-specific stem cells suggests that they may respond differently to DNA damage than
downstream cells. In this issue of Cell Stem Cell, two groups address this hypothesis by examining DNA
damage responses in hematopoietic stem and progenitor cells (Milyavsky et al., 2010; Mohrin et al., 2010).

Nearly 50 years after Till and McCulloch hematopoietic cell radiation sensitivity stem cell (HSC) radiosensitivity are
discovered clonal myeloerythroid cells (Till and McCulloch, 1961), the mecha- beginning to be unraveled. In contrast
in the pursuit of developing assays for nisms and the meaning of hematopoietic to somatic cells that are frequently

Cell Stem Cell 7, August 6, 2010 ª2010 Elsevier Inc. 145


postmitotic and/or short-lived, tissue-
specific stem cells must function
throughout life in order to mediate
ongoing tissue homeostasis and repair.
The enormous functional demands and
longevity of stem cells suggests that
stem cells, particularly those from highly
regenerative compartments such as the
skin and blood, may be equipped with
effective DNA damage response (DDR)
mechanisms to ensure genomic integrity
over a lifetime. With this said, it is also
increasingly clear that HSCs specifically
accumulate DNA damage during aging,
a mechanism believed to contribute to
the elevated incidence of cancer in the
elderly (Rossi et al., 2007).
In the hematopoietic system, only
HSCs, memory T, and memory B lympho-
cytes can self-renew; the hierarchical
differentiation scheme underlying HSC
lineage commitment makes DNA damage
accrual in HSCs a dangerous prospect.
In all stem cell-driven tissues, misrepaired
lesions arising in stem cells can be propa-
gated both to daughter stem cells and
downstream progeny through the pro-
cesses of self-renewal and differentiation.
In such a way, mutations arising in stem
cells can thus be potentiated throughout
all cells of the tissue (Rossi et al., 2008).
Given that the cost of mutation accumula-
tion in stem cells is so high, it has been
postulated that stem cells may be
uniquely privileged with properties that
either limit DNA damage accrual or effec-
tively respond to it once damage is
incurred. Consistent with this model,
stem cells from diverse tissues possess
a variety of cytoprotective properties.
For example, stem cells from a variety of
tissues exhibit elevated ABC transporter
activity, resulting in an increased ability
to pump genotoxic compounds out of
the cell. Stem cells from several adult
tissues reside in a noncycling, quiescent
posture, minimizing the chance of rep-
lication error (Yamazaki et al., 2006).
Moreover, quiescent stem cells are meta-
bolically inactive and as such, generate
low levels of endogenous free radicals
and reactive oxygen species (Tothova
et al., 2007).
Less is known, however, about how
stem cells respond to DNA lesions once
they have happened. Overexpression of
Bcl2 provides relative protection of
HSCs in terms of cell death responses
(Domen and Weissman, 2003), but it is
146 Cell Stem Cell 7, August 6, 2010 ª2010 Elsevier Inc.
unclear whether the rescued cells convert
DNA damage to mutations and/or translo-
cations. These important issues have now
been addressed in two papers published
in this issue of Cell Stem Cell (Milyavsky
et al., 2010; Mohrin et al., 2010). The
studies used the hematopoietic stem
and progenitor compartments of mice
(Mohrin et al., 2010) and humans (Milyav-
sky et al., 2010) to address the mecha-
nisms through which stem and progenitor
cells respond to DNA double-strand
breaks (DSBs) induced by ionizing
radiation.
In the mice, Mohrin et al. addressed this
issue by using hematopoietic stem and
progenitor cells (HSPCs) and myeloid pro-
genitors isolated from young mice. They
found that at certain doses of ionizing irra-
diation (2 Gy), HSPCs were more resistant
than downstream myeloid progenitors,
a result consistent with earlier studies
(Meijne et al., 1991). In the absence of
ATM, all populations show equal radiation
sensitivity, suggesting that ATM is
involved in the differential DDR of HSPCs
versus progenitors. This pathway induces
both cell-cycle arrest and expression of
prosurvival genes in HSPCs, followed by
DNA repair. In contrast, damaged myeloid
progenitors tended to be eliminated by
apoptosis. The fact that quiescent HSPCs
are almost uniformly in the G0 phase of
the cell cycle suggested that they would
be obliged to use the error-prone nonho-
mologous end joining (NHEJ) pathway,
in contrast to the more high fidelity homol-
ogous recombination (HR) pathway to
repair DSBs. This hypothesis was tested
by immunostaining and with a functional
assay, which confirmed that HSPCs pref-
erentially utilized NHEJ in contrast to
cycling progenitor cells. HSPCs driven
into cycle prior to damage, however,
enabled access to HR to repair DSBs.
Interestingly, HSPCs driven into cycle still
exhibited radioresistance over myeloid
progenitors. The reliance of quiescent
HSPCs on the error-prone NHEJ pathway
raised the possibility that DNA damage
might be preferentially misrepaired in
these cells. To test this point, the authors
irradiated cycling or quiescent HSPCs
and then examined genomic integrity in
their progeny by spectral karyotyping.
Strikingly, irradiated quiescent HSPCs
gave rise to progeny that harbored chro-
mosomal abnormalities at a higher fre-
quency and in higher numbers than
Cell Stem Cell
Previews
when HSPCs were prestimulated into
cycle. Taken together, these findings sug-
gest that mouse HSC quiescence and reli-
ance on NHEJ may be an important
mechanism contributing to mutagenesis
at the stem cell level that may serve as
the origin of certain blood cell cancers.
In a complementary study published in
this issue, Milyavsky et al. addressed
DDR by using hematopoietic stem and
progenitor cells isolated from human
cord blood (CB). Strikingly, and in con-
trast to the murine study, irradiated
human CB HSPCs exhibit a slower DNA
damage repair kinetic and proved less
radio-resistant than downstream pro-
genitor cells. Knockdown of p53, a domi-
nant form of p53, or ectopic expression
of BCL2 revealed that the p53-BCL2
pathway was involved in mediating the
apoptotic response of irradiated cord
blood progenitors. Knockdown and func-
tional evaluation of ASPP1, a molecular
regulator of p53-dependent apoptosis,
suggested that stem cell radiosensitivity
was at least in part mediated by ASPP1.
To further evaluate the functional conse-
quence of DNA damage and response
on the function of CB HSPCs, the authors
assessed their viability in the NOD/SCID
repopulation assay. Irradiation (3 Gy)
quantitatively abrogated the ability of
cord blood cells to repopulate recipients,
which could be partially rescued by
knocking down p53 or overexpressing
BCL2. The authors then examined the
affect of p53 and BCL2 on genomic integ-
rity during serial transplantation. Interest-
ingly, DNA damage (as measured by
gH2AX foci) levels were similar in primitive
hematopoietic progenitors expressing
either p53 or BCL2 in primary transplant
recipients, yet in secondary recipients,
p53 knockdown alone led to increased
indication of DNA damage accrual, which
correlated with a diminished capacity of
the p53-deficient cells to repopulate the
secondary recipients. On the basis of
these findings, the authors propose that
the p53 apoptotic response is uncoupled
from DNA damage accumulation and
suggest an apoptosis-independent role
for p53 in human CB HSC self-renewal.
Collectively, the findings of Mohrin and
Milyavsky and their respective colleagues
have opened the door for a deeper
understanding of DDR in stem cells. Strik-
ingly, a number of findings in these
studies suggest that murine and human
Cell Stem Cell
Previews
hematopoietic stem and progenitor
compartments may respond to DNA in
different ways. For example, the DNA
damage repair kinetics of human CB
HSPCs was delayed compared to down-
stream progenitors, in contrast to murine
HSPCs, which exhibited a faster repair
kinetic. Similarly, whereas human HSPCs
were proapoptotic and more radiosensi-
tive than downstream progenitors, murine
HSPCs were more radioresistant ap-
peared more antiapoptotic than progen-
itor cells. Interestingly, a recent paper by
Blanpain and colleagues examining the
DNA damage response in murine hair
follicle stem cells reported apoptotic and
repair kinetic properties more similar to
murine HSPCs, suggesting that species-
specific differences may underlie these
different findings (Sotiropoulou et al.,
2010).
Perhaps the most interesting possibility
is that DDR differences seen in CB HSPCs
from human versus bone marrow or
mobilized HSPCs from young mice relate
to the different anatomical locations, or
to the stage of ontogeny from which
the studied cells were derived, given
that that both anatomical location and
ontogeny are known to significantly
impact HSC function (Rossi et al., 2008).
Because many hematopoietic disorders
that stem from DNA damage accrual arise
during aging, these results also stress the
importance of examining DNA damage
response and damage accrual during
ontogeny and aging.
ACKNOWLEDGMENTS
I.W. owns significant stock in Amgen, Inc., from
past service on its SAB. He is a director and
consultant and holds stock in Stem Cells, Inc.,
and he is a former director and consultant and
stockholder with Cellerant, Inc. He has consulted
for Fate Therapeutics.
REFERENCES
Domen, J., and Weissman, I.L. (2003). Exp. Hema-
tol. 31, 631–639.
Meijne, E.I., van der Winden-van Groenewegen, R.
J., Ploemacher, R.E., Vos, O., David, J.A., and
Huiskamp, R. (1991). Exp. Hematol. 19, 617–623.
Cell Stem Cell 7, August 6, 2010 ª2010 Elsevier Inc. 147
Milyavsky, M., Gan, O.I., Trottier, M., Komosa, M.,
Tabach, O., Notta, F., Lechman, E., Hermans, K.
G., Eppert, K., Konovalova, Z., et al. (2010). Cell
Stem Cell 7, this issue, 186–197.
Mohrin, M., Bourke, E., Alexander, D., Warr, M.R.,
Barry-Holson, K., Le Beau, M.M., Morrison, C.G.,
and Passegué, E. (2010). Cell Stem Cell 7, this
issue, 174–185.
Rossi, D.J., Bryder, D., Seita, J., Nussenzweig, A.,
Hoeijmakers, J., and Weissman, I.L. (2007). Nature
447, 725–729.
Rossi, D.J., Jamieson, C.H., and Weissman, I.L.
(2008). Cell 132, 681–696.
Sotiropoulou, P.A., Candi, A., Mascre, G.,
De Clercq, S., Youssef, K.K., Lapouge, G., Dahl,
E., Semeraro, C., Denecker, G., Marine, J.C.,
et al. (2010). Nat. Cell Biol. 12, 572–582.
Till, J.E., and McCulloch, C.E. (1961). Radiat. Res.
14, 213–222.
Tothova, Z., Kollipara, R., Huntly, B.J., Lee, B.H.,
Castrillon, D.H., Cullen, D.E., McDowell, E.P.,
Lazo-Kallanian, S., Williams, I.R., Sears, C., et al.
(2007). Cell 128, 325–339.
Yamazaki, S., Iwama, A., Takayanagi, S., Morita,
Y., Eto, K., Ema, H., and Nakauchi, H. (2006).
EMBO J. 25, 3515–3523.