**3.3 Association of the SDF1-3'A allele with a good mobilizing capacity**

As the clinicians have defined mobilization failure as <2x106 CD34+ cells/kg obtained within 4 apheresis days, two mainly group of patients emerged: the subjects with a good capacity of mobilization who collected ≥2x106 CD34+ cells/kg obtained within 4 apheresis days. Others with a poor mobilizing capacity and didn't collect 2x106 CD34+ cells/kg within 4 apheresis days. For the healthy allogenic PBSC donors, the mobilization failure was defined as <3x106 CD34+ cells/kg obtained within 4 apheresis days.

When considering the SDF1-3'A polymorphism, significant difference was observed in the SDF1-3'A allele carriers and GG carriers (p=0.023). A higher concentration of CD34+ cells in the leukapheresis products was detected in SDF1-3'A positive patients compared to GG homozygous subjects

Besides, a lower increase in the GG genotypes was observed in the "poor" mobilizer group compared to the "good" ones reaching a statistical significance (p=0.023; OR =0.494; CI (95%) [0.268-0.912]) (Table 4).

Thus, the SDF1-3'A allele carriers, especially the SDF1-3'AA homozygous individuals in the group of healthy allogenic PBSC donors had a better mobilization potential (table 4).


Abbreviations: OR, odds ratio; af, allele frequency; gf, genotype frequency; CI, confidence interval (CI=95%); Corrected *p* value; NS, not significant; \*, for SDF-1 polymorphism; \*\* for MMP-9 polymorphism, Good mobilizers (>2X106 CD34/kg), Poor mobilizers (<2x106CD34/kg)

Table 4. Allele and genotype frequencies of *SDF-1,* and *MMP-*9 polymorphisms in mobilized peripheral blood patients and healthy controls

In this table are provided:

308 Advances in Hematopoietic Stem Cell Research

In table 3 are provided: all genotypic and allelic frequencies according to each polymorphism studied and corresponding to all patients. Distribution of genotypic and allelic frequencies by each disease included in this study. Then, all frequencies are calculated by statistical software

For the group of NHL, the distribution of the SDF1-3'A polymorphism was significantly different between patients and healthy controls especially for the A allele which seemed to be associated to this disease (p=0,019). Moreover, a decrease in GG genotype frequency compared to the control group was observed too reaching a statistically significance

Concerning the MMP-9 C-1562T polymorphism, like the MM group, high significant differences were seen especially for the T allele (P<0.05; OR=4.055; CI (95%) [1.901-8.646]) and CT genotypes (P<0.05; OR=6.333; CI (95%) [2.754-14.567]). Similar results were obtained concerning the distribution of the MMP-9 C-1562T polymorphism in the group of Hodgkin's disease where significant differences were found in the T allele and CT genotype

While, the distribution of the SDF1-3'A polymorphism was not significantly different between the group of patients with AML and the control group, MMP-9 C-1562T distribution was significantly different essentially for the T allele (p=0.019, OR= 7.298, CI (95%) [1.511-35.249])

When considering the GNB3 C825T polymorphism, we observed that the TT genotype was more frequent in patient with MM and NHL with respectively 20.69% and 15.15% compared to the Hodgkin's disease group (only 10.52%). Whereas, the CC genotype was more frequent

As the clinicians have defined mobilization failure as <2x106 CD34+ cells/kg obtained within 4 apheresis days, two mainly group of patients emerged: the subjects with a good capacity of mobilization who collected ≥2x106 CD34+ cells/kg obtained within 4 apheresis days. Others with a poor mobilizing capacity and didn't collect 2x106 CD34+ cells/kg within 4 apheresis days. For the healthy allogenic PBSC donors, the mobilization failure was

When considering the SDF1-3'A polymorphism, significant difference was observed in the SDF1-3'A allele carriers and GG carriers (p=0.023). A higher concentration of CD34+ cells in the leukapheresis products was detected in SDF1-3'A positive patients compared to GG

Besides, a lower increase in the GG genotypes was observed in the "poor" mobilizer group compared to the "good" ones reaching a statistical significance (p=0.023; OR =0.494; CI

Thus, the SDF1-3'A allele carriers, especially the SDF1-3'AA homozygous individuals in the

group of healthy allogenic PBSC donors had a better mobilization potential (table 4).

and the CT genotypes (p=0.004, OR= 12.444, CI (95%) [2.485-62.319]) Table 3. So the presence of the MMP-9 C-1562T might be associated with this disease.

**3.3 Association of the SDF1-3'A allele with a good mobilizing capacity** 

defined as <3x106 CD34+ cells/kg obtained within 4 apheresis days.

SPSS 16.0 as well as p value and odd ratios (OR) are provided.

(p=0.029).

frequencies (p<0.05; Table 3).

in the NHL group (24.24%) (Table 3).

homozygous subjects

(95%) [0.268-0.912]) (Table 4).

All genotypic Allelic frequencies designed as "gf" and allelic frequencies designed as "af" of SDF1-3'A and MMP-9 C-1562T polymorphisms in all the study populations (all patients), then in a group of healthy blood donors (as control group)

Then when, dividing the whole patients according to their mobilization capacity into: good mobilizers (>2X106 CD34/kg), and poor mobilizers (<2x106CD34/kg).

OR designed as odd radio and p value of all genotypic and allelic frequencies are provided in the table by using statistical software (SPSS 16.0) as it was mentioned above in section materials and methods-statistical analysis.

However, when considering the group of remobilizers in our study population we have observed that 48% of subjects were GG, 12% were AA and 40% were GA. This led us to consider a probable association of the GG genotypes to mobilization failure.

Distribution of SDF1-3'A, GNB3 C825T and MMP-9 C-1562T Polymorphisms

fair candidate gene variants to these 3 hematological diseases.

investigators [De Oliveira KB et al, 2009; Rabkin CS et al, 1999].

transplantation by using a PCR-RFLP analysis.

Marilyn Kozak, 2004; Gavin S. Wilkie et al, 2003].

Hodgkin's lymphoma [De Oliveira KB et al, 2009].

representation of AML [A Zafiropoulos et al, 2004].

of patients (15 patients).

suggest the possible role of such variant in the pathogenesis of NHL.

**4. Discussion** 

in HSC CD34+ from Peripheral Blood of Patients with Hematological Malignancies 311

In the present study, we investigated the effect of polymorphisms in the genes SDF-1, GNB3 and MMP-9 on the outcome of mobilization of peripheral blood stem cells for autologous

We observed a significant association for SDF-1 and MMP-9 polymorphisms exclusively in patients with MM, NHL and Hodgkin's disease suggesting that these polymorphisms are

In fact, Association of these polymorphisms to cancer has been previously reported by many

Our results were in agreement with other studies suggesting that SDF1-3'A polymorphism is a genetic determinant of NHL [Gabriela Gonçavales de Olivera Cavassin et al, 2004]. Furthermore; as the SDF1-3'A polymorphism is situated in the mRNAs of 3'UTR region (untranslated region) which has been identified as an important regulator of the mRNA transcript, as well as the translated product [Catia Andreassi and Antonella Riccio, 2004;

The second polymorphism studied encoded for MMP-9, J. Arai et al, have reported that SDF-1 mRNAs abundantly expressed in stromal cells from the lymph nodes of patients with malignant lymphoma, so that 3'A carriers NHL are good candidates for presenting proliferation of neoplasic cells in the lymph nodes since that SDF-1 variant is associated with an increase of

De Oliveira KB et al, when studying distribution of SDF1-3'A polymorphism have reported also a significant difference in genotype distribution between NHL patients (GG: 51.4%; GA: 47.1%; AA: 1.5%) compared to healthy controls (GG: 65.6%; GA: 28.9%; AA: 5.5%). Whereas, they didn't find any significant differences in genotypes distributions with breast cancer and

Moreover, previous reports on AIDS related non-Hodgkin's lymphoma (NHL) demonstrated that the CXCL12–3'A chemokine variant was associated with approximate doubling of the NHL risk in heterozygotes and an approximately fourfold increase in homozygotes [Rabkin CS et al, 1999; A Zafiropoulos et al, 2004]. Hence, this might let us

In this present work, we did not find a significant association between SDF1-3'A polymorphism and our group of patients with AML, this could be due to the lower number

However, Dommange et al, have reported the implication of SDF1-3'A polymorphism in the clinical representation of acute myeloid leukemia in 86 patients with AML, as an association between this polymorphism and the risk of tissue infiltration by malignant cell was established by an increased release of the blast from the bone marrow in the blood in the SDF1-3'A carriers suggesting that this SDF-1 variant is associated with clinical

MMP-9 is a zinc-dependent proteinase, which is involved in numerous physiological and pathological processes. In the present study, we reported the distribution of the functional

SDF-1 levels [J. Arai et al, 2000; Gabriela Gonçavales de Olivera Cavassin et al, 2004].

For the MMP-9 C-1562T polymorphism, significant difference was obtained with CT genotypes between the two groups (p=0.004; OR= 0.297; CI (95%) [0.125-0.703]).

For the GNB3 C825T polymorphism, we didn't observe any difference between the 2 groups of poor and good mobilizers.

This let us consider that there's no association between GNB3 C825T polymorphism and the capacity of mobilization of hematopoietic stem cells.

For the group of healthy PBSC donors, and with respect to our classification according to mobilization failure (<3x106 CD34/kg within 4 apheresis days), we have found an important association of SDF1-3'A distribution with higher mobilization yield of hematopoietic stem cells CD34+ reaching a higher statistical significance (p=0.001; OR=12.6; table 5).

Besides, we have observed a similar increase in the SDF1-3'G allele in the intermediate to poor mobilizers' subgroup reaching a statistical significance (p=0.035; OR=1.25; table 4). Similarly, the association was already observed when comparing the genotypic frequencies between the two subgroups.

The AA genotype was absent in the poor mobilizer subgroup, then was highly increased in the other subgroup reaching a statistical significance (p=0.035; OR=1.25).


While, the GG genotype was more represented in the poor mobilizers and the differences were significant too (p=0.001; OR=0.079; table 4).

Abbreviations: OR, odds ratio; af, allele frequency; gf, genotype frequency; CI, confidence interval (CI=95%); Corrected *p* value; NS, not significant; \*, for SDF-1 polymorphism; \*\* for MMP-9 polymorphism, for healthy allogenic PBPC donors: Good mobilizers (>3X106 CD34/kg), Poor mobilizers (<3x106CD34/kg)

Table 5. Allele and genotype frequencies of *SDF-1,* and *MMP-9* polymorphisms in mobilized peripheral blood of healthy allogenic PBSC donors
