- Email: [email protected]
P17 Sperm and lymphocyte aneuploidy in patients with translocations
S28
10th International Congress on Preimplantation Genetic Diagnosis
translocation to rule out possibility of germ cell mosaicism or chimerism. In the case of presence of normal sperms, PGD can be offered to these couples. This study is very important in which it will change genetic counseling attitude towards this type of translocation carriers. P16 Preimplantation Genetic Diagnosis (PGD) cycles for translocations carriers. Results in a public hospital O. L´ opez1 , O. Martinez-Pasarell1 , E. García-Guix´ e2 , M. Sandalinas2 , A. Polo3 , A. Garcia1 , A. Mata1 , P. Viscasillas3 , o Puigvert; 2 Reprogenetics L. Bassas1 . 1 Embryology Lab, Fundaci´ Spain; 3 Gynecology, Hospital de Sant Pau, Barcelona, Spain Objective: Carriers of chromosomal rearrangements have and increased risk for pregnancy losses and repeated failures in IVF cycles. Couples with balanced translocations are usually good candidates for PGD. We have analyzed the results of the embryology laboratory in our public hospital. Material and Methods: Data were collected from March 2001 until February 2010. A total of 28 IVF-PGD cycles were performed. Reciprocal translocations (RecT) were present in 19 cycles (15 of female carriers and 4 of male carriers) and Robertsonian translocation (RobT) in 9 cycles (4 of female carriers and 5 of male carriers). Female age was 34.9±2.5 (mean±SD). A total of 204 embryos were biopsied on day 3 and analyzed for rearranged chromosomes by FISH. Aneuploidy screening was also performed in selected cases. Normal/ balanced embryos were transferred in day 4 or 5. Data obtained in IVF-PGD cycles were analyzed and compared (RecT vs. RobT; male vs. female carriers). Results: No significant differences were observed in the total number of oocytes retrieved, microinjected oocytes and total obtained embryos regarding the type of translocation (RecT vs. RobT). However, for female carriers we obtained more embryos in cycles of RecT than RobT (8.7±3.8 vs. 6.7±1.2; p = 0.03). The total number of unbalanced embryos was higher for RecT than for RobT (5.4±2.4 vs. 3.0±1.3; p = 0.02). This difference was also observed for female carriers (p = 0.04). Seven out the twentyeight IVF-PGD cycles (1 male-RobT; 1 female-RobT; 1 maleRecT and 4 female-RecT) did not have balanced/normal embryos for transfer. In a female-RecT case, transfer was cancelled because normal/balanced embryos had other chromosomal abnormalities. The mean number of normal/balanced embryos transferred was 1.3±1.0 and no statistical differences were observed regarding the type of translocation, nor the sex carrier. Pregnancy rates were slightly better in RobT cycles than in RecT cycles (33.3% vs. 21.0% per cycle; 42.9% vs. 30.7% per transfer), but differences were not significant. Conclusions: Carriers of reciprocal translocation carriers have less favorable prognosis than those with Robertsonian translocations. We have observed more unbalanced embryos in cycles of reciprocal translocations, mainly in female carriers. P17 Sperm and lymphocyte aneuploidy in patients with translocations N. Zotova, E. Markova, I. Timofeeva, O. Kazantseva, V. Artukhova, A. Svetlakov. Krasnoyarsk Center for Reproductive Medicine, P.O. Box 2714, Krasnoyarsk, 660037, Russia Aim: PGD patients with translocations are considered to be at high risk of translocated chromosome missegregation and aneuploidy because of interchromosomal effect (ICE). So far, ICE has not been well investigated. We performed aneuploidy screening for chromosomes participating in translocation and for five additional chromosomes (13, 18, 21, X, and Y) in sperm and lymphocytes for patients with translocations. Method: In this study, we analyzed 17 patients with translocations and reproductive failure (aged 34.87±1.68) and 10 healthy
men of control group selected according to WHO-criteria for sperm donors (aged 32.00±2.28). Sperm cells were available only for five patients with translocations. Eleven patients balanced and two had Robertsonian translocations, four unbalanced translocations. Standard cytogenetic investigation was performed for all men. We used 5-coloure FISH with “MultiVysion PGT” probes “Abbott Molecular Inc.” (13, 18, 21, X, Y) for sperm cells and lymphocytes. Additionally we analyzed chromosomes participating in translocation: 2, 8, 9, 10, 14, 15, and 22. One thousand blood and sperm cells per person were scored. Results: We did not find any ICE in blood cells of patients with translocations. Sex chromosome aneuploidy level was 0.41±0.04%, similar as in the control group: 0.30±0.03%. There were no differences in aneuploidy levels of the 13th, 18th, and 21st chromosomes. The rate of normal/balanced sperm of five translocation carriers was 89% (4452/5000). The rate of normal sperm of ten control men was 98.8% (9883/10000). ICE in sperm was found in one patient with t(13;14): increased XYdisomy level and disomy 18; and in another patient with t(13;22): increased sex chromosome aneuloidy and diploidy level. So, our conclusion was that meiotic mistakes of chromosome segregation took place. In lymphocytes of 9 patients and in sperm of 5 patients we found a missegregation of chromosomes participating in translocation. Conclusion: We found the ICE in sperm cells of some patients with translocation and did not find in lymphocytes. So, we recommended to perform sperm FISH before PGD for translocation patients. phone/fax +7(391)2640895, e-mail: [email protected]
P18 PGD for a woman carrier of a balanced reciprocal translocation (11;22) and a man with high expression of 16q22 fragile site O. Martinez-Pasarell1 , O. Lopez1 , E. Garcia-Guix´ e2 , M. Sandalinas2 , A. Polo3 , A. Mata1 , A. Garcia1 , P. Viscasillas3 , L. Bassas1 . 1 Embryology Laboratory, Fundacio Puigvert; 2 Reprogenetics Spain; 3 Gynecology, Hospital de Sant Pau, Barcelona, Spain Introduction: Carriers of balanced reciprocal translocations are at increased risk of spontaneous abortions due to the high frequency of unbalanced gametes (20 80% depending specific chromosomes). Fragile sites appear as gaps or chromosome breaks in some of the metaphases from a given individual. The 16q22 fragile site (FRA16B) is not associated with any gene and no specific clinical problems have been reported for carriers. However, some fragile sites are related with chromosome instability. The aim of this study is to describe the outcome of the PGD cycle for a couple with a balanced translocation (11;22) and FRA16B. Materials and Methods: A 34 year-old woman was carrier of 46,XX,t(11;22)(q23;q11.2) karyotype. Her partner (normozoospermic, 35 year-old) had normal karyotype (46,XY) with FRA16B expression in 50% of metaphases analysed. They presented 2-year history of subfertility with a clinical abortion a biochemical pregnancy. FISH sperm analysis for 13, 18, 21, X and Y chromosomes was performed. Meiotic segregation of chromosome 16 (Tel 16pter/Tel 16 qter) was also analysed with the aim to discard 16q segregation problems. An IVFPGD cycle was conducted to select normal/balanced embryos for chromosomes 11 and 22. Probable gains or losses due to 16q22 fragile site were also considered. In order to detect interchromosomal effects, chromosomes X, Y, 13, 14, 18 and 21 were included in the PGD cycle.
10th International Congress on Preimplantation Genetic Diagnosis
translocation to rule out possibility of germ cell mosaicism or chimerism. In the case of presence of normal sperms, PGD can be offered to these couples. This study is very important in which it will change genetic counseling attitude towards this type of translocation carriers. P16 Preimplantation Genetic Diagnosis (PGD) cycles for translocations carriers. Results in a public hospital O. L´ opez1 , O. Martinez-Pasarell1 , E. García-Guix´ e2 , M. Sandalinas2 , A. Polo3 , A. Garcia1 , A. Mata1 , P. Viscasillas3 , o Puigvert; 2 Reprogenetics L. Bassas1 . 1 Embryology Lab, Fundaci´ Spain; 3 Gynecology, Hospital de Sant Pau, Barcelona, Spain Objective: Carriers of chromosomal rearrangements have and increased risk for pregnancy losses and repeated failures in IVF cycles. Couples with balanced translocations are usually good candidates for PGD. We have analyzed the results of the embryology laboratory in our public hospital. Material and Methods: Data were collected from March 2001 until February 2010. A total of 28 IVF-PGD cycles were performed. Reciprocal translocations (RecT) were present in 19 cycles (15 of female carriers and 4 of male carriers) and Robertsonian translocation (RobT) in 9 cycles (4 of female carriers and 5 of male carriers). Female age was 34.9±2.5 (mean±SD). A total of 204 embryos were biopsied on day 3 and analyzed for rearranged chromosomes by FISH. Aneuploidy screening was also performed in selected cases. Normal/ balanced embryos were transferred in day 4 or 5. Data obtained in IVF-PGD cycles were analyzed and compared (RecT vs. RobT; male vs. female carriers). Results: No significant differences were observed in the total number of oocytes retrieved, microinjected oocytes and total obtained embryos regarding the type of translocation (RecT vs. RobT). However, for female carriers we obtained more embryos in cycles of RecT than RobT (8.7±3.8 vs. 6.7±1.2; p = 0.03). The total number of unbalanced embryos was higher for RecT than for RobT (5.4±2.4 vs. 3.0±1.3; p = 0.02). This difference was also observed for female carriers (p = 0.04). Seven out the twentyeight IVF-PGD cycles (1 male-RobT; 1 female-RobT; 1 maleRecT and 4 female-RecT) did not have balanced/normal embryos for transfer. In a female-RecT case, transfer was cancelled because normal/balanced embryos had other chromosomal abnormalities. The mean number of normal/balanced embryos transferred was 1.3±1.0 and no statistical differences were observed regarding the type of translocation, nor the sex carrier. Pregnancy rates were slightly better in RobT cycles than in RecT cycles (33.3% vs. 21.0% per cycle; 42.9% vs. 30.7% per transfer), but differences were not significant. Conclusions: Carriers of reciprocal translocation carriers have less favorable prognosis than those with Robertsonian translocations. We have observed more unbalanced embryos in cycles of reciprocal translocations, mainly in female carriers. P17 Sperm and lymphocyte aneuploidy in patients with translocations N. Zotova, E. Markova, I. Timofeeva, O. Kazantseva, V. Artukhova, A. Svetlakov. Krasnoyarsk Center for Reproductive Medicine, P.O. Box 2714, Krasnoyarsk, 660037, Russia Aim: PGD patients with translocations are considered to be at high risk of translocated chromosome missegregation and aneuploidy because of interchromosomal effect (ICE). So far, ICE has not been well investigated. We performed aneuploidy screening for chromosomes participating in translocation and for five additional chromosomes (13, 18, 21, X, and Y) in sperm and lymphocytes for patients with translocations. Method: In this study, we analyzed 17 patients with translocations and reproductive failure (aged 34.87±1.68) and 10 healthy
men of control group selected according to WHO-criteria for sperm donors (aged 32.00±2.28). Sperm cells were available only for five patients with translocations. Eleven patients balanced and two had Robertsonian translocations, four unbalanced translocations. Standard cytogenetic investigation was performed for all men. We used 5-coloure FISH with “MultiVysion PGT” probes “Abbott Molecular Inc.” (13, 18, 21, X, Y) for sperm cells and lymphocytes. Additionally we analyzed chromosomes participating in translocation: 2, 8, 9, 10, 14, 15, and 22. One thousand blood and sperm cells per person were scored. Results: We did not find any ICE in blood cells of patients with translocations. Sex chromosome aneuploidy level was 0.41±0.04%, similar as in the control group: 0.30±0.03%. There were no differences in aneuploidy levels of the 13th, 18th, and 21st chromosomes. The rate of normal/balanced sperm of five translocation carriers was 89% (4452/5000). The rate of normal sperm of ten control men was 98.8% (9883/10000). ICE in sperm was found in one patient with t(13;14): increased XYdisomy level and disomy 18; and in another patient with t(13;22): increased sex chromosome aneuloidy and diploidy level. So, our conclusion was that meiotic mistakes of chromosome segregation took place. In lymphocytes of 9 patients and in sperm of 5 patients we found a missegregation of chromosomes participating in translocation. Conclusion: We found the ICE in sperm cells of some patients with translocation and did not find in lymphocytes. So, we recommended to perform sperm FISH before PGD for translocation patients. phone/fax +7(391)2640895, e-mail: [email protected]
P18 PGD for a woman carrier of a balanced reciprocal translocation (11;22) and a man with high expression of 16q22 fragile site O. Martinez-Pasarell1 , O. Lopez1 , E. Garcia-Guix´ e2 , M. Sandalinas2 , A. Polo3 , A. Mata1 , A. Garcia1 , P. Viscasillas3 , L. Bassas1 . 1 Embryology Laboratory, Fundacio Puigvert; 2 Reprogenetics Spain; 3 Gynecology, Hospital de Sant Pau, Barcelona, Spain Introduction: Carriers of balanced reciprocal translocations are at increased risk of spontaneous abortions due to the high frequency of unbalanced gametes (20 80% depending specific chromosomes). Fragile sites appear as gaps or chromosome breaks in some of the metaphases from a given individual. The 16q22 fragile site (FRA16B) is not associated with any gene and no specific clinical problems have been reported for carriers. However, some fragile sites are related with chromosome instability. The aim of this study is to describe the outcome of the PGD cycle for a couple with a balanced translocation (11;22) and FRA16B. Materials and Methods: A 34 year-old woman was carrier of 46,XX,t(11;22)(q23;q11.2) karyotype. Her partner (normozoospermic, 35 year-old) had normal karyotype (46,XY) with FRA16B expression in 50% of metaphases analysed. They presented 2-year history of subfertility with a clinical abortion a biochemical pregnancy. FISH sperm analysis for 13, 18, 21, X and Y chromosomes was performed. Meiotic segregation of chromosome 16 (Tel 16pter/Tel 16 qter) was also analysed with the aim to discard 16q segregation problems. An IVFPGD cycle was conducted to select normal/balanced embryos for chromosomes 11 and 22. Probable gains or losses due to 16q22 fragile site were also considered. In order to detect interchromosomal effects, chromosomes X, Y, 13, 14, 18 and 21 were included in the PGD cycle.