**3. Results**

#### **3.1 Patient's distributions according to their CD34+ cell yield and failure rates**

Overall 83% of patients included in this study collected ≥2x106 CD34+ cells/kg after a maximum of 4 aphereses, among them 20% collected 2-5x106 CD34cells/kg, and 63% collected ≥5x106 CD34 cells/kg. Beside, 10% are remobilizers as they did not achieve the threshold of CD34+ cell yield of 2x106 CD34/kg within 4 apheresis days and are subjects to another mobilization protocol. Among them, the group of NHL represented the highest rate (40%), the lower ones, the group of MM and AML, which represented respectively 19% and 13%. By contrast, others are designed as first mobilizers (90%) since they have already collected ≥2x106CD34+ cells/kg after a maximum of 4 aphereses days. Amongst them the group of multiple myeloma was the most frequent (40%), thereafter the group of Non-Hodgkin's lymphoma (34%) and Hodgkin's disease with 26%. For the patients included in this study, mobilization failure was defined as <2x106 CD34+ cells/kg obtained within 4 apheresis days. So, especially MM patients collected ≥5x106CD34+ cells/kg and contained the highest CD34+ cell yield (8,89x106 CD34/kg for MM, and 5,51x106 CD34/kg for the others patients). Furthermore, the fact that MM patients had higher yield of CD34+ cells compared to NHL and HD is likely since that NHL and HL patients are frequently more heavily pretreated with cytotoxic chemotherapy than patients with MM [Iskra Pusic et al, 2008] (figure 4).

#### **3.2 Analysis of the studied polymorphisms in the 4 subgroups of patients according to disease: A comparison between healthy donors of PBSC and patients**

According to this study, SDF1-3'A and MMP-9 C-1562 T polymorphisms were significantly different between the patients and healthy controls (table 3). Particularly, we found significant differences in all the allelic and genotypic frequencies of the SDF1-3'A polymorphism in the MM group (p<0.05; OR=3.245 CI (95%) [1.830-5.753] for A allele; p= 0.017; OR= 3.324 CI (95%) [1.182-9.348]; p= 0.009; OR= 2.072 CI (95%) [1.200-3.580] for AA and GA genotypes, respectively).

Concerning the MMP-9 C-1562 T polymorphism its distribution was significantly different in the same MM group of patients compared to the control group, significant differences were observed exclusively for the T allele (p=0.041; OR=2.295 CI (95%) [1.020-5.168]) and also for the CC and CT genotypes (p= 0.039; p= 0.004; Table 3).

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

in HSC CD34+ from Peripheral Blood of Patients with Hematological Malignancies 307

Table 3. Allele and genotype frequencies of *SDF-1, GNB3* and *MMP-9 polymorphisms* in the

groups of patients with MM, NHL, Hodgkin's' Disease and AML

A number of first mobilization and remobilization in database Distribution of remobilizers in the 4 subgroup of patients

Distribution of good and poor mobilizers of PBPC CD34+ in the study population and by sex

Fig. 4. Overview of autologous stem cell transplantation database by disease as well as the distribution of good/poor mobilizers of PBSC CD34+ within the study population and by sex is already represented


Table 3. Allele and genotype frequencies of *SDF-1, GNB3* and *MMP-9 polymorphisms* in the groups of patients with MM, NHL, Hodgkin's' Disease and AML

306 Advances in Hematopoietic Stem Cell Research

A number of first mobilization and remobilization in database Distribution of remobilizers in

Distribution of good and poor mobilizers of PBPC CD34+ in the study population and by sex

Fig. 4. Overview of autologous stem cell transplantation database by disease as well as the distribution of good/poor mobilizers of PBSC CD34+ within the study population and by

sex is already represented

NHL 40%

the 4 subgroup of patients

19% HD

28%

AML 13% MM

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

in HSC CD34+ from Peripheral Blood of Patients with Hematological Malignancies 309

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)

peripheral blood patients and healthy controls

materials and methods-statistical analysis.

then in a group of healthy blood donors (as control group)

mobilizers (>2X106 CD34/kg), and poor mobilizers (<2x106CD34/kg).

consider a probable association of the GG genotypes to mobilization failure.

In this table are provided:

Table 4. Allele and genotype frequencies of *SDF-1,* and *MMP-*9 polymorphisms in mobilized

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 when, dividing the whole patients according to their mobilization capacity into: good

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

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

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

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 (p=0.029).

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 frequencies (p<0.05; Table 3).

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]) 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.

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