6位罹患Y染色体长臂无精区C段显微缺失并无精症患者的临床描述及利用睪丸取精并单一精虫卵细胞质内注射技术所获致的结果
6位罹患Y染色体长臂无精区C段显微缺失并无精症患者的临床描述及利用睪丸取精并单一精虫卵细胞质内注射技术所获致的结果
Clinical characterization of 6 azoospermic men with microdeletion of the AZFc region of Y chromosome, and the outcome of intracytoplasmic injection of sperm obtained by testicular sperm extraction from these men
江俊明、林娟如 － 健安妇产科不孕症中心
Chun-Ming Chiang, Chuan-Ju Lin －Jiahn-An OB/GYN Clinic and Infertility Center, Keelung
Introduction:
AZFc, the most frequently deleted region of the Y chromosome in infertile males, is a de-novo microdeletion found in ~1: 4000 males (13% of azoospermic men and ~6% of men with severe oligospermia). In this study we accumulated six azoospermic men with a Y chromosome microdeletion confined to the AZFc region, and we report their clinical characterization and outcome of ICSI using sperm obtained by TESE from these men.
In a study by Van Golde et al (Hum Reprod 2001;16) reported there were significant differences between severely oligospermic patients with AZFc deletions and the severely oligospermic men with intact Y-DNA in outcome of ICSI using ejaculated sperm in term of fertilization rate (55% VS,71%; 55%:AZFc deleted group, 71% intact Y-DNA group; P<0.001) and embryo quality (lower embryo quality in AZFc deletions group).
In our study, we also compared the outcome of TESE/ICSI in patients with AZFc deletions to that of non-obstructive men with intact Y-DNA to discern whether there were any significant differences in fertilization, implantation, clinical pregnancy and live delivery rate achieved by using testicular sperm for ICSI.
Materials:
Six couples suffering primary infertility in which the male partner were azoospermia, caused by microdeletion of the AZFc region of the Y chromosome, participated in this study. For these 6 male partner, their history, physical examination, karyotype, FSH, LH, testosterone, inhibin B, and testis histology were retrospectively accumulated in our clinical practice. All of the women underwent controlled ovarian stimulation. Open testis biopsies for TESE were conducted 1 day prior to or on the day of oocyte retrieval. Mature sperm, extracted from the biopsied tissues in a wet preparation were used for ICSI. The outcome of ICSI was evaluated from the fertilization, embryo-cleavage rate, implantation rate, clinical pregnancy rate and somatic status of resultant babies. The outcome of TESE/ICSI also were compared to that of non-obstructively azoospermic men with intact Y-DNA in term of fertilization rate, implantation rate, clinical pregnancy rate and live delivery rate. Y-DNA analysis was done in Chiang’s andrological laboratory (Chiang HS, et al. Asian J Androl 2004; 6)
Results:
The clinical characteristics of AZFc deleted men and its outcome of TESE/ICSI and the comparison of outcome of TESE/ICSI between the studied group with AZFc deletions with control group with intact Y-DNA are showed in table I, II, and III, respectively.
Conclusion：
The six azoospermic men with Y-AZFc deletions were healthy individual with no major illness. Penile anatomy was normal in all men and no patient showed any overt sign of decreased virilization. Both of average testosterone and LH values were in normal limits. Therefore, genes in the AZFc region do not appear to impair interstitial Leyding cell function.
The serum FSH levels in six studied men were slightly higher compared to that in normal population implies that Sertolic-germ cell function was compromised in azoospermic patients with AZFc deletions.
The variance in histopathological findings showed in our studies' men with Y-AZFc deletions might be the consequence of being modified by each man's particular environment and genetic background (Krausz C, et al. Int. J. Androl 2003;26).
In our study, 7 out of 8 TESE attempts could find sperm implied that absences of genes in Y-AZFc region may not smother spermatogenesis entirely. Our study demonstrated that although the absence of genes in Y-AZFc region quantitatively reduces sperm density so severely that the infertility and sterility are the rules and natural procreation the exception, the sperm almost could be found by TESE in these azoospermic men with AZFc deletions. It perhaps due to the existence of autosomal homologues for most of the transcriptional units that are found in the Y-AZFc region (Kuroda-Kauaquchi T, et al. Nat Gent 2001;29).
Our data also provided encouragement for infertile men with AZFc deletions who are azoospermic – it is very likely that sperm can be harvested from testis tissue, thereby providing an opportunity for biological fatherhood.
Our study showed that sperm quality of azoospermic men with AZFc deletions is inferior to that of non-obstructively azoospermic patients with intact Y-DNA in term of fertilization rate, implantation rate and clinical pregnancy rate. Our data also had the trend of lower live delivery rate in couples with AZFc deletions compared to the control group with intact Y-DNA.
Table I. Clinical characteristics of Y-AZFc deleted men
|
Case |
Age at diagnosis (years) |
Medical history |
Testis biopsy prior to this visit |
FSH mIU/ml |
LH mIU/ml |
Testosterone ng/dl |
Inhibin-B pg/ ml |
Sperm found by ESP |
Histology at Biopsy / TESE |
Sperm found by TESE |
|
WangTP1a |
36 |
- |
Yes |
7.31 |
7.08 |
1.92 |
- |
- |
SCOb |
found |
|
WangTP2 |
38 |
- |
- |
- |
- |
- |
- |
- |
- |
found |
|
WangTP3 |
38 |
- |
- |
- |
- |
- |
- |
- |
- |
found |
|
Wang SA |
30 |
- |
- |
5.42 |
9.0 |
5.01 |
167.8 |
Found (very few) |
SCO/MA |
not found |
|
Lee JW |
30 |
- |
Yes |
17.7 |
14.0 |
4.75 |
150.3 |
- |
SCO |
found |
|
Cheng JC |
38 |
Varicocele, Post-operation |
- |
29.7 |
11.5 |
4.33 |
- |
- |
- |
found |
|
Lee HT |
28 |
- |
Yes |
11.5 |
3.89 |
8.12 |
112.9 |
- |
MAc |
found |
|
Cheng IH |
37 |
- |
Yes |
5.85 |
3.05 |
5.28 |
129.9 |
- |
SCO |
found |
|
Mean±SD |
33.2±4.31 |
- |
- |
12.92±9.42 (normal range 2-10) |
8.09±4.28 (normal range <25) |
4.90±1.99 (normal range 2.7-10.7) |
140.23±23.91 (normal range 93-327) |
- |
- |
- |
|
a: Wang TP received 3 TESE attempt in our clinic. b: SCO = Sertoli cell-only c: MA = maturation arrest All of the Y-AZFc deleted men are normal 46,XY karyotype. |
||||||||||
Table II. Outcome of ICSI
|
Male partner |
Female partner age (years) |
Injected oocytes |
2PN |
Cleaved embryos |
Embryos Transferred |
Gestational sacs |
outcome |
|
WangTP1 |
32 |
11 |
6 |
6 |
6 |
0 |
0 |
|
WangTP2 |
34 |
11 |
7 |
7 |
7 |
0 |
0 |
|
WangTP3 |
34 |
10 |
4 |
4 |
4 |
0 |
0 |
|
Wang SA |
23 |
none |
- |
- |
- |
- |
- |
|
Lee JW |
29 |
7 |
2 |
2 |
2 |
0 |
0 |
|
Cheng JC |
34 |
14 |
2 |
2 |
2 |
0 |
0 |
|
Lee HT |
27 |
11 |
3 |
3 |
3 |
0 |
0 |
|
Cheng IH |
36 |
11 |
8 |
8 |
8 |
2 |
1 boy 1 girl |
Table III. TESE/ICSI outcomes of non-obstructively azoospermic men with Y-AZFc deletion compared with that of non-obstructively azoospermic men with intact Y-DNA (control group)
|
|
AZFc deleted |
Non-obstructive |
P |
|
No. of cycles |
8 |
28 |
|
|
No. of patients |
6 |
27 |
|
|
Female age(y), mean±SD |
31.1 ± 4.4 |
30.0 ± 3.8 |
0.3093a |
|
Sperm obtained / TESE attempts, % (n) |
87.5 (7/8) |
50.0 (14/28) |
0.1038b |
|
2PN / injected oocytes, % (n) |
42.7 (32/75) |
58.1 (86/148) |
0.0413c |
|
No. of embryos transferred, mean±SD |
4.6 ± 2.4 |
5.9 ± 2.3 |
|
|
Implantation rate ( No. of gestational sacs/ No. of transferred embryos), %(n) |
6.3 (2/32) |
31.2 (24/77) |
0.0113c |
|
Clinical pregnancy rate (No. of clinical pregnancies / No. of embryo transfer cycles), %(n) |
14.3 (1/7) |
76.9 (10/13) |
0.0166b |
|
Live delivery rate (No. of live delivery / No. of embryo transfer cycles), %(n) |
14.3 (1/7) |
61.5 (8/13) |
0.0703b |
a：Wilcoxon Two-Sample Test
b：Fisher’s Exact Test
c：Chi-Square Test