• HSCs contribute to neutrophil recovery.

  • HSCs symmetrically divide, forming clonal pools.

Hematopoietic stem cells (HSCs) are assumed to be rare, infrequently dividing, long-lived cells not involved in immediate recovery after transplantation. Here, we performed unprecedented high-density clonal tracking in nonhuman primates and found long-term persisting HSC clones to actively contribute during early neutrophil recovery, and to be the main source of blood production as early as 50 days after transplantation. Most surprisingly, we observed a rapid decline in the number of unique HSC clones, while persisting HSCs expanded, undergoing symmetric divisions to create identical siblings and formed clonal pools ex vivo as well as in vivo. In contrast to the currently assumed model of hematopoietic reconstitution, we provide evidence for contribution of HSCs in short-term recovery as well as symmetric expansion of individual clones into pools. These findings provide novel insights into HSC biology, informing the design of HSC transplantation and gene therapy studies.

1.
Sawai
CM
,
Babovic
S
,
Upadhaya
S
, et al
.
Hematopoietic stem cells are the major source of multilineage hematopoiesis in adult animals
.
Immunity
.
2016
;
45
(
3
):
597
-
609
.
2.
Morcos
MNF
,
Zerjatke
T
,
Glauche
I
, et al
.
Continuous mitotic activity of primitive hematopoietic stem cells in adult mice
.
J Exp Med
.
2020
;
217
(
6
):
e20191284
.
3.
Chapple
RH
,
Tseng
YJ
,
Hu
T
, et al
.
Lineage tracing of murine adult hematopoietic stem cells reveals active contribution to steady-state hematopoiesis
.
Blood Adv
.
2018
;
2
(
11
):
1220
-
1228
.
4.
Matsuzaki
Y
,
Kinjo
K
,
Mulligan
RC
,
Okano
H
.
Unexpectedly efficient homing capacity of purified murine hematopoietic stem cells
.
Immunity
.
2004
;
20
(
1
):
87
-
93
.
5.
Notta
F
,
Doulatov
S
,
Laurenti
E
,
Poeppl
A
,
Jurisica
I
,
Dick
JE
.
Isolation of single human hematopoietic stem cells capable of long-term multilineage engraftment
.
Science
.
2011
;
333
(
6039
):
218
-
221
.
6.
Yamamoto
R
,
Wilkinson
AC
,
Ooehara
J
, et al
.
Large-scale clonal analysis resolves aging of the mouse hematopoietic stem cell compartment
.
Cell Stem Cell
.
2018
;
22
(
4
):
600
-
607.e4
.
7.
Wilson
NK
,
Kent
DG
,
Buettner
F
, et al
.
Combined single-cell functional and gene expression analysis resolves heterogeneity within stem cell populations
.
Cell Stem Cell
.
2015
;
16
(
6
):
712
-
724
.
8.
Biasco
L
,
Pellin
D
,
Scala
S
, et al
.
In vivo tracking of human hematopoiesis reveals patterns of clonal dynamics during early and steady-state reconstitution phases
.
Cell Stem Cell
.
2016
;
19
(
1
):
107
-
119
.
9.
Six
E
,
Guilloux
A
,
Denis
A
, et al
.
Clonal tracking in gene therapy patients reveals a diversity of human hematopoietic differentiation programs
.
Blood
.
2020
;
135
(
15
):
1219
-
1231
.
10.
Scala
S
,
Basso-Ricci
L
,
Dionisio
F
, et al
.
Dynamics of genetically engineered hematopoietic stem and progenitor cells after autologous transplantation in humans
.
Nat Med
.
2018
;
24
(
11
):
1683
-
1690
.
11.
Adair
JE
,
Enstrom
MR
,
Haworth
KG
, et al
.
DNA barcoding in nonhuman primates reveals important limitations in retrovirus integration site analysis
.
Mol Ther Methods Clin Dev
.
2020
;
17
:
796
-
809
.
12.
Beard
BC
,
Adair
JE
,
Trobridge
GD
,
Kiem
HP
.
High-throughput genomic mapping of vector integration sites in gene therapy studies
.
Methods Mol Biol
.
2014
;
1185
:
321
-
344
.
13.
Radtke
S
,
Adair
JE
,
Giese
MA
, et al
.
A distinct hematopoietic stem cell population for rapid multilineage engraftment in nonhuman primates
.
Sci Transl Med
.
2017
;
9
(
414
):
eaan1145
.
14.
Radtke
S
,
Pande
D
,
Cui
M
, et al
.
Purification of human CD34+CD90+ HSCs reduces target cell population and improves lentiviral transduction for gene therapy
.
Mol Ther Methods Clin Dev
.
2020
;
18
:
679
-
691
.
15.
Biddy
BA
,
Kong
W
,
Kamimoto
K
, et al
.
Single-cell mapping of lineage and identity in direct reprogramming
.
Nature
.
2018
;
564
(
7735
):
219
-
224
.
16.
Jones
RJ
,
Celano
P
,
Sharkis
SJ
,
Sensenbrenner
LL
.
Two phases of engraftment established by serial bone marrow transplantation in mice
.
Blood
.
1989
;
73
(
2
):
397
-
401
.
17.
Kim
S
,
Kim
N
,
Presson
AP
, et al
.
Dynamics of HSPC repopulation in nonhuman primates revealed by a decade-long clonal-tracking study
.
Cell Stem Cell
.
2014
;
14
(
4
):
473
-
485
.
18.
Wu
C
,
Li
B
,
Lu
R
, et al
.
Clonal tracking of rhesus macaque hematopoiesis highlights a distinct lineage origin for natural killer cells
.
Cell Stem Cell
.
2014
;
14
(
4
):
486
-
499
.
19.
Bystrykh
LV
,
Verovskaya
E
,
Zwart
E
,
Broekhuis
M
,
de Haan
G
.
Counting stem cells: methodological constraints
.
Nat Methods
.
2012
;
9
(
6
):
567
-
574
.
20.
Sun
J
,
Ramos
A
,
Chapman
B
, et al
.
Clonal dynamics of native haematopoiesis
.
Nature
.
2014
;
514
(
7522
):
322
-
327
.
21.
Bernitz
JM
,
Kim
HS
,
MacArthur
B
,
Sieburg
H
,
Moore
K
.
Hematopoietic stem cells count and remember self-renewal divisions
.
Cell
.
2016
;
167
(
5
):
1296
-
1309.e10
.
22.
Upadhaya
S
,
Sawai
CM
,
Papalexi
E
, et al
.
Kinetics of adult hematopoietic stem cell differentiation in vivo
.
J Exp Med
.
2018
;
215
(
11
):
2815
-
2832
.
23.
Mahmud
N
,
Devine
SM
,
Weller
KP
, et al
.
The relative quiescence of hematopoietic stem cells in nonhuman primates
.
Blood
.
2001
;
97
(
10
):
3061
-
3068
.
24.
Abkowitz
JL
,
Catlin
SN
,
Guttorp
P
.
Evidence that hematopoiesis may be a stochastic process in vivo
.
Nat Med
.
1996
;
2
(
2
):
190
-
197
.
25.
Abkowitz
JL
,
Golinelli
D
,
Harrison
DE
,
Guttorp
P
.
In vivo kinetics of murine hemopoietic stem cells
.
Blood
.
2000
;
96
(
10
):
3399
-
3405
.
26.
Catlin
SN
,
Guttorp
P
,
Abkowitz
JL
.
The kinetics of clonal dominance in myeloproliferative disorders
.
Blood
.
2005
;
106
(
8
):
2688
-
2692
.
27.
Shepherd
BE
,
Kiem
HP
,
Lansdorp
PM
, et al
.
Hematopoietic stem-cell behavior in nonhuman primates
.
Blood
.
2007
;
110
(
6
):
1806
-
1813
.
28.
Ashcroft
P
,
Manz
MG
,
Bonhoeffer
S
.
Clonal dominance and transplantation dynamics in hematopoietic stem cell compartments
.
PLoS Comput Biol
.
2017
;
13
(
10
):
e1005803
.
29.
Park
DS
,
Akuffo
AA
,
Muench
DE
, et al
.
Clonal hematopoiesis of indeterminate potential and its impact on patient trajectories after stem cell transplantation
.
PLoS Comput Biol
.
2019
;
15
(
4
):
e1006913
.
30.
Kiel
MJ
,
Yilmaz
OH
,
Iwashita
T
,
Yilmaz
OH
,
Terhorst
C
,
Morrison
SJ
.
SLAM family receptors distinguish hematopoietic stem and progenitor cells and reveal endothelial niches for stem cells
.
Cell
.
2005
;
121
(
7
):
1109
-
1121
.
31.
Radtke
S
,
Pande
D
,
Cui
M
, et al
.
Sort-purification of human CD34+CD90+ cells reduces target cell population and improves lentiviral transduction
.
bioRxiv
.
2020
.
32.
Velten
L
,
Haas
SF
,
Raffel
S
, et al
.
Human haematopoietic stem cell lineage commitment is a continuous process
.
Nat Cell Biol
.
2017
;
19
(
4
):
271
-
281
.
33.
Chang
KC
,
Wang
C
,
Wang
H
.
Balancing self-renewal and differentiation by asymmetric division: insights from brain tumor suppressors in Drosophila neural stem cells
.
Bioessays
.
2012
;
34
(
4
):
301
-
310
.
34.
Sugiarto
S
,
Persson
AI
,
Munoz
EG
, et al
.
Asymmetry-defective oligodendrocyte progenitors are glioma precursors
.
Cancer Cell
.
2011
;
20
(
3
):
328
-
340
.
35.
Homem
CCF
,
Knoblich
JA
.
Drosophila neuroblasts: a model for stem cell biology
.
Development
.
2012
;
139
(
23
):
4297
-
4310
.
36.
Cicalese
A
,
Bonizzi
G
,
Pasi
CE
, et al
.
The tumor suppressor p53 regulates polarity of self-renewing divisions in mammary stem cells
.
Cell
.
2009
;
138
(
6
):
1083
-
1095
.
37.
Wu
M
,
Kwon
HY
,
Rattis
F
, et al
.
Imaging hematopoietic precursor division in real time
.
Cell Stem Cell
.
2007
;
1
(
5
):
541
-
554
.
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