Stromal elements and Engraftment
in Autologous Hematopoetic
Pi
Progen t
itor Cellll Transpl
t
an for
Myeloma
Muthu Veeraputhiran, Zeba Singh, Bart
Barlogie and Michele Cottler-
Cottler Fox
UAMS/MIRT
Little Rock, Arkansas
Background
Subset of myeloma patients with
with delayed
engraftment
No explanation for delayed ENG
Need for HPC boost
Objective
Correlate engraftment (ENG) with
with :
Stromal parameters
Disease parameters
Engraftment p
gparameters
WBC Engraftment
Engraftment
·
ANC > 500/cu.mm
Platelet Engraftment
·
> 20,000/cu.mm
·
> 50 000/
,
cu.mm
Disease parameters
Duration of disease
Prior transplant
transplant
Cd
Con itii
ditioning chemoth
therapy for transplant
Total viable CD34+ cells infused
Stromal variables
Fat content vs hematopoetic cellularity ( %)
Regenerating adipocytes (+, -)
Fat Necrosis (+, -)
Hematopoiesis (Endosteal vs sinusoidal or both) (+, -)
Bone remodelling (osteoblasts vs osteoclasts or both)
(+, -)
Reticulin fibrosis ( 0,1,2,3,4)
Stromal vascularity ( +, -)
Stromal damage ( +, -)
- Stromal edema
- Granular degeneration
- Stromal hemorrhage
Anatomy of Stroma
Stromal cells
cells
- fibroblasts or reticular cells
- adipocytes
Stromal support system
- progenitor/stem cells
(hematopoetic, mesenchymal, endothelial
prog
pgenitors)
- macrophages, endothelial cells
- Extracellular matrix
- fibrous proteins, glycoproteins and
proteoglycans
The Osteoblastic and Vascular Niches in Bone
* Adapted from www.Biology-online.org
Ot
Ost
b
eo l
blast
t
ac ii
tiv t
ity
Osteoclast activity
Stromal vascularity
Increased vascularity
- Dilated sinusoids, increased number of
sinusoids
- Complex network of capillaries
Dilated sinusoids
Rt
Regenera i
ting d
a i
dipocytes
Signs of Stromal Inj
gjury
Stromal edema: clear fluid-like space
space
surrounding stromal cells in extracellular
matrix (ECM)
()
Granular degeneration: PAS
PA +
S material in the
stroma
Stromal hemorrhage: extravasation of RBCs
around sinusoids into ECM
Strom
o al
a edem
ede a
Granular degeneration
Stromal hemorrhage
Marrow Fibrosis (MF)
()
Grading of Fibrosis5
g
- 0: No reticulin
- 1+: Occasional fine fibers + foci of fine fiber
network
- 2+: Fine fiber network throughout most of the
section arranged
arranged in parallel; no coarse fibers
- 3+: Diffuse, fiber network with scattered thick,
coarse, criss-crossing fibers,
g, but no true collagen
g
(- trichrome stain)
- 4+: Diffuse, often coarse, fiber network with areas
fl
of co lll
i
agen
t
za i
tion (+ tri h
c rome t
s i
a )
n
-5. 0 to 1+: Normal;
Bauermeister D et al. Am 2+
J
Borderline;
Clin Pathol. 1971;
3+
56:
to
24-31 4+: Severe
Reticulin fibrosis
Gi' Sil
H & E
Gomori's Silver
Masson's Trichrome
Materials and Methods
Two groups
- Normal engrafters
-
( PLT ENG to 20,000 and 50,000 < 2 SD, n=26)
- Poor engrafters
-
( PLT ENG to 20,000/50,000 > 2 SD off mean, n=12)
Exclusion criteria
-
No ENG data
-
Infections
Staining
-
H & E stain
-
Gomori's silver and Masson's Trichrome stains
* SD = Standard deviation
Statistical analyses
Wald
l chi-
chi square test
test for
for logistic regression
-
Odds ratio
Cox regression
regression model
-
Hazards ratio
-
Ui
Uni
it
variate regression analysis
-
Multivariate regression analysis
Comparison of Normal and Poor engrafters
ENGRAFTMENT
NORMAL
POOR
ENGRAFTERS
ENGRAFTERS
Mean
Median
Mean
Median p-value
ANC 500
12
12
18
15
0.002
PLT 20000
13
13
46
46
< 0.001
PLT 50000
16
16
56
57
< 0.001
Stromal Variables with Positive Correlation with ENG
VARIABLES
ASSOCIATION
ANALYSIS
Endosteal
WBC and PLT ENG
Univariate
hematopoesis
Multivariate
Stromal vascularity
PLT ENG
Univariate
Osteoclastic activity
WBC ENG
Univariate
Multivariate
Osteoblastic activity
activity
WBC ENG
ENG and PLT
PL to
to
Univariate
20000
* p-value < 0.05 is considered significant
Results
No correlation of engraftment with:
- Fibrosis (increased odds to be seen in poor
ENG)
- Adipocyte content vs hematopoetic cellularity
- Stromal hemorrhage
- Stromal damage
Acknowledgements
Dr. Michele Cottler-
Cottler Fox
Dr. Zeba Singh
DB
Dr. Bart Barlogie
Ms. Jackie Szymonifka ( CRAB, Seattle, WA)